Introduction to Medical Technology purposes: sales (medical representative), researcher, teacher, pre-med Medical techs role: in the labbehind

the scenes. Only contact with the patients is during !"#$O%OM&. SECTIONS OF THE LABORATORY: ! Clinical Che"i#try: analytical monitoring o' (cleanse blood by removal o' e<cess 'luid, chemical parameters resulting 'rom minerals, and wastes) or +idney physiological or biochemical processes in the transplant (surgical procedure to implant a body. healthy +idney into a patient with +idney $lood chemistry- sugar(glucose, disease or +idney 'ailure) with the patient. cholesterol(triglycerides, en)ymes LABORATORY TESTS *omponents o' $lood: =4ormal values 'or many tests are determined by the patient8s age and gender. ,e'erence ac+ed ,ed $lood *ells values can also vary by laboratory latelet *oncentrate Routine urinaly#i#: 6imple > cheap -resh--ro)en lasma: screening test. O'ten the 'irst test *ryoprecipitate- treatment o' hemophilia conducted i' +idney problems are . suspected. 6ample is e<amined physically: *ryosupernatant- treatment o' color, odor, appearance, and concentration /hemophilia $ (speci'ic gravity)7 chemically: protein, 01234#& -54*%2O4 %#6%6: variety o' glucose, > p!7 and microscopically: individual tests and procedures to evaluate presence o' cellular elements (,$*s, how well +idneys are 'unctioning. . doctor who ?$*s, and epithelial cells), bacteria, orders 1-%s and uses the results to assess the crystals, and casts (structures 'ormed by 'unctioning o' the +idneys is called a the deposit o' protein, cells, and other nephrologist. substances in the +idneys8s tubules). %he +idneys: natural 'iltration system7 per'orm 2' results @ abnormal, /A o' the 'ollowing vital 'unctions: removing metabolic waste additional tests is usually per'ormed to products 'rom the bloodstream, regulating the pinpoint the cause and the level o' decline body8s water balance, and maintaining the p! in +idney 'unction. o' the body8s 'luids. 9/: ;uarts o' Creatinine clearance te#t. #''iciency as blood(minute are circulated through the the +idneys clear creatinine 'rom the +idneys, where waste chemicals are 'iltered blood. *reatinine, a waste product o' out and eliminated 'rom the body (along with muscle energy metabolism, is produced at e<cess water) in the 'orm o' urine. 1idney a constant rate that is proportional to the 'unction tests help to determine i' the +idneys individual8s muscle mass. .ll creatinine are per'orming their tas+s ade;uately. 'iltered by the +idneys in a given amount =4eed *OM "#%# !26%O,& to evaluate the o' time is e<creted in the urine, ma+ing patient8s 'ood and drug inta+e. rescription creatinine clearance a very speci'ic and over-the-counter medications, as well as measurement o' +idney 'unction. %he test some 'ood and beverages, can a''ect blood is per'ormed on a timed urine specimena and urine 1-% results. cumulative sample collected over a two to Many conditions can a''ect the ability o' the BC-hour period. 3etermination o' the blood +idneys. *onditions cause either an acute creatinine level is also re;uired to calculate decline or a chronic decline in 'unction. $oth the urine clearance. can result in a build-up o' to<ic waste $rea clearance te#t. 5rea is a waste substances in the blood. *linical laboratory tests that measure the levels o' substances product that is created by protein normally regulated by the +idneys can metabolism and e<creted in the urine. %he determine the cause and e<tent o' +idney urea clearance test re;uires a blood dys'unction. 5se: 5rine > $lood. sample measures amount o' urea in the %he nephrologist uses these results in a bloodstream and two urine specimens, number o' ways. Once a diagnosis is made (one hour apart,) to determine the amount that +idney disease is present and what +ind o' urea that is 'iltered or cleared by the o' +idney disease is causing the problem, the +idneys into the urine. nephrologist may recommend a speci'ic %est compares the level o' creatinine in treatment. .lthough there is no speci'ic drug urine with the creatinine in the blood and is therapy that will prevent the progression o' used to estimate the glomerular 'iltration +idney disease, the doctor will ma+e rate (D-,). -or a BC-hour urine collection, recommendations 'or treatment to slow the normal results are EFG/HE m"(min 'or adult disease as much as possible. -or instance, the males younger than CF, and IFG/BJ doctor might prescribe blood pressure m"(min 'or adult 'emales younger than CF. medications, or treatments 'or patients with -or people over CF, values decrease by K.J diabetes. 2' +idney disease is getting worse, m"(min 'or each decade o' li'e. the nephrologist may discuss hemodialysis

 $rine o#"olality te#t. Measure dissolved particles in urine. More precise than speci'ic gravity to evaluate the ability o' the +idneys to concentrate or dilute the urine. 4ormal +idneys@ e<crete water as 'luid inta+e. 2' 'luid inta+e, the +idneys e<crete water. 6ample may be: 'irst thing in the morning, multiple timed samples, or a cumulative sample collected over a BChour period. %ypically, patient @ highprotein diet A drin+ no 'luids the night be'ore. ?ith restricted 'luid inta+e (concentration testing), osmolality should be greater than IFF mOsm(+g o' water. ?ith increased 'luid inta+e (dilution testing), osmolality should be less than /FF mO6m(+g in at least one o' the specimens collected. . BChour urine osmolality should average HFFG EFF mOsm(+g. . random urine osmolality should average JFFGIFF mOsm(+g. $rine %rotein te#t. !ealthy +idneys 'ilter all proteins 'rom the bloodstream and then reabsorb them, allowing no to slight amounts o' protein into the urine. ersistent presence o' signi'icant amounts o' protein in the urine is an important indicator o' +idney disease. . positive screening test 'or protein (included in a routine urinalysis ) on a random urine sample is usually 'ollowed up with a test on a BC-hour urine sample that more precisely measures the ;uantity o' protein. . BC-hour urine collection should contain no more than /JF mg o' protein. $rine #odiu". . BC-hour urine sodium should be within LJGBFF mmol(day. %here are also several blood tests that can aid in evaluating +idney 'unction. %hese include: Blood urea nitrogen te#t &B$N'! 5rea @ byproduct o' protein metabolism. -ormed in the liver, this waste product is then 'iltered 'rom the blood and e<creted in the urine by the +idneys. %he $54 test measures the amount o' nitrogen contained in the urea. !igh $54 levels can indicate +idney dys'unction, but because $54 is also a''ected by protein inta+e and liver 'unction, the test is usually done together with a blood creatinine, a more speci'ic indicator o' +idney 'unction. $lood urea nitrogen ($54) should average IGBF mg(d". Creatinine te#t. %his test measures blood levels o' creatinine, a by-product o' muscle energy metabolism that, similar to urea, is 'iltered 'rom the blood by the +idneys and e<creted into the urine. roduction o' creatinine depends on an person8s muscle mass, which usually 'luctuates very little. ?ith normal +idney 'unction, then, the amount o' creatinine in the blood remains

relatively constant and normal. -or this reason, and because creatinine is a''ected very little by liver 'unction, an elevated blood creatinine level is a more sensitive indicator o' impaired +idney 'unction than the $54. *reatinine should be F.IG/.B mg(d" 'or males, and F.KGF.E mg(d" 'or 'emales. Other (lood te#t#. Measurement o' the blood levels o' other elements regulated in part by the +idneys can also be use'ul in evaluating +idney 'unction. %hese include sodium, potassium, chloride, bicarbonate, calcium, magnesium, phosphorus, protein, uric acid, and glucose. 5ric acid levels 'or males should be H.JG L.B mg(d" and 'or 'emales B.KGK.F mg(d". = $54, alone, is suggestive, but not diagnostic 'or +idney dys'unction as it can be a''ected by other 'actors. .bnormally plasma creatinine is a more speci'ic indicator o' +idney disease than $54. =creatinine clearance values and urea @ability o' +idneys to 'ilter these waste products 'rom the blood and e<crete them. .s clearance levels, blood levels o' creatinine, uric acid and urea . =2nability o' +idneys to concentrate the urine a'ter 'luid inta+e, or dilute the urine a'ter 'luid inta+e during osmolality testing@+idney 'unction. = ersistent presence o' protein, in amounts that e<ceed the normal BC-hour urine value, usually indicates some type o' +idney disease. =6ome +idney problems are the result o' another disease process, such as diabetes or hypertension. 3octors should in'orm patients about how their disease or its treatment will a''ect +idney 'unction, as well as the di''erent measures patients can ta+e to help prevent these changes. )*.,32.* -54*%2O4 %#6%6 Creatine *ina#e &C*': %otal *17 *reatine phospho+inase7 * 1 Related te#t#: *1-M$7 Myoglobin7 %roponin7 *ardiac biomar+ers #n)yme that stores high energy creatine phosphate (mostly in muscle cells.) 2t is widely distributed in tissue, with highest activities 'ound in s+eletal muscle, heart muscle and brain tissue. *1 is present in tissue sources, including the bladder, placenta, gastrointestinal tract, thyroid, uterus, +idney, lung, prostate, spleen, liver and pancreas. H di''erent 'orms o' *1 in the body7 isoen)ymes: *1-MM ('ound in your s+eletal muscles and heart), *1-M$ ('ound mostly in your heart), *1-$$ ('ound mostly in your brain).

%he small amount o' *1 that is normally in the blood comes mainly 'rom your muscles. %he *1 in your brain almost never gets into the blood. $lood sample ta+en 'rom arm. =$lood levels o' *1@muscle or heart cells are inMured. *hest pain or symptoms o' heart attac+, concentration o' *1 in blood (C-K hours a'ter heart attac+). 2t reaches its highest level in /I to BC hours and returns to normal within B to H days. =amount o' *1 in blood when s+eletal muscles are damaged. En+y"e ,unction: cataly)e the conversion o' creatine to phosphocreatine by applying itsel' in the consumption o' adenosine triphosphate, the generation o' adenosine diphosphate, and the reverse reaction. .denosine triphosphate is a vital source o' energy in biochemical reactions li+e in the s+eletal muscle, brain, and smooth muscle or all tissues. ?hile the phosphocreatine acts as an energy reservoir 'or the ;uic+ regeneration o' adenosine triphosphate. =4ormal conditions@ very little creatine +inase circulating in the blood o' the average, healthy human being. %he test speci'ically measures the blood levels o' certain muscle and brain en)yme proteins, thus, the normal result 'or 'emales ranges between /F G LE units per liter (5(") and /L G /CI 5(" in males. . lower than normally low level o' creatine +inase shows that you have been drin+ing e<cessively. .lcohol liver disease and rheumatoid arthritis are two o' the most common possibilities that e<ist with respect to lowered levels o' creatine +inase. =2' the test reveals that the level o' creatine +inase circulating in the blood is higher than it should be in normal conditions, then chances are that the human body in ;uestion has su''ered damage either to the muscle or the brain. 2n 'act, astronomical levels o' creatine +inase are indicative o' inMuries, rhabdodomyolysis, myocardial in'arction, myocarditis, myositis, malignant hypethermia, Mc"eod syndrome, neuroleptic malignant syndrome, and hypothyroidism. Lactate -ehydrogena#e &L-H' #n)yme that cataly)es the interconversion o' lactic and pyruvic acids. 2t is a hydrogentrans'er en)yme that uses the coen)yme 4.3A. "3 is widely distributed in the body. activities are 'ound in the heart, liver, s+eletal muscle, +idney, and erythrocytes7 lesser amounts are 'ound in the lung, smooth muscle, and brain.

"3! @ general indicator o' e<istence and severity o' acute or chronic tissue damage and, sometimes, as a monitor o' progressive conditions such as some cancers, +idney disease, and liver disease. 2t used to be ordered to help diagnose and monitor a heart attac+, but the troponin test has largely replaced "3! in this role. "3! isoen)ymes may be used in di''erential diagnosis to help determine which organs are li+ely to be involved. En+y"e ,unction: screening test when a doctor suspects cellular or tissue damage. 2' the total "3!, he may order "3! isoen)ymes or other tests such as ."%, .6% or ." to help diagnose the condition and to help determine which organs are involved. Once the acute or chronic problem is diagnosed, total "3! levels may be used at regular intervals to monitor its progress and(or resolution. "3! may ordered to monitor damage caused by muscle trauma or inMury and to help identi'y hemolytic anemia. !emolytic anemia is caused by the brea+age o' red blood cells, either because they are unusually 'ragile or because something is mechanically brea+ing them, such as an arti'icial heart valve. "3! and "3! isoen)ymes may still be occasionally ordered along with *1 and *1M$ when someone has symptoms o' a heart attac+, but this is increasingly rare. 2n most cases today, the doctor will order troponin levels along with the *1 and *1M$ instead o' "3!. er'ormed to evaluate the presence o' tissue damage. 2t is an intracellular en)yme 'ound in almost all body tissues and is released a'ter there is tissue damage. %he highest concentrations are 'ound in the heart, liver, lungs, +idneys, red blood cells and s+eletal muscle cells. .'ter damaged to tissues 'rom trauma, ischemia, or acid(base imbalance, "3! is released into the bloodstream. %he results o' this test indicate that tissue damage has occurred but cannot pinpoint the speci'ic location o' damage. ?hen total "3 is elevated to at least /HF 25(", the test "actate dehydrogenase isoen)ymes$lood should be per'ormed to narrow down the source o' tissue damage. "3! 'unctions as a catalyst the interconversion o' pyruvate and lactate. %he active e<ercising muscles convert (and red blood cells metaboli)e) glucose to lactate. "actate released into the blood is ta+en up by the liver which converts lactate bac+ to glucose and releases glucose into the blood. "3! test is per'ormed as a prognostic 'actor in colorectal carcinoma and is being studied 'or its use as a prognostic 'actor in

myelodysplastic syndromes, with higher levels being associated with shorter survival . 4ormal ,esult @/CF-BIF 5(": "3! levels typically will rise as the cellular destruction begins, pea+ a'ter some time period, and then begin to 'all. 2' someone has a heart attac+, blood levels o' total "3! will rise within BC to CI hours, pea+ in B to H days, and return to normal in /F to /C days. !igher-than-normal levels may indicate: $lood 'low de'iciency (ischemia) *erebrovascular accident (such as a stro+e) !eart attac+ !emolytic anemia 2n'ectious mononucleosis "iver disease (e<ample: hepatitis) "ow blood pressure Muscle inMury Muscular dystrophy 4ew abnormal tissue 'ormation (usually cancer) ancreatitis %issue death ?ith some chronic and progressive conditions, and some drugs, moderately "3! levels may persist. "ow and normal levels o' "3! do not usually indicate a problem. levels sometimes @ ingests large amounts o' ascorbic acid (vitamin *). A#%artate A"inotran#,era#e&AST': 6erum glutamic-o<aloacetic transaminase(6DO%)7 .spartate transaminase7 .6%(."% ratio Related te#t#: ."%7 ." 7 DD%7 $ilirubin7 "iver panel7 .lbumin7 %otal rotein #n)yme belonging to the class o' trans'erases7 commonly re'erred to as a transaminase ('acilitate certain chemical reactions within cells) and is involved in the trans'er o' an amino group between aspartate and +eto acids. -ound in amounts in heart muscle, liver and muscle cells7 and amounts in other tissues such as in the +idneys, pancreas and erythrocytes. En+y"e ,unction: = Myocardial 2n'arction(M2). G .6% can assist in determining the timing and e<tent o' a recent M2 ("ess speci'ic than creatine phospho+inase (* 1), *1M$, myglobin, troponins, and lactic dehydrogenase ("3!).) = .6% determination valuable in these disorders > diseases: acute pancreatitis, muscle disease, trauma, severe burn, and in'ectious mononucleosis. =.6% @ aid in the diagnosis o' liver disease7 not speci'ic 'or liver disease, but combine with other en)ymes to monitor course o' various liver disorders.

.mount o' .6% / number o' cells a''ected by the disease or inMury7 level o' elevation depends on length o' time that blood is tested a'ter inMury. =6erum .6% levels: elevated I hours a'ter cell inMury , pea+ at BC-HK hours, return to normal in three to seven days. =2' cellular inMury is chronic, .6% levels will remain elevated. =2' no 'urther cardiac inMury occurs: .6% level rises within K-/F hours a'ter an acute attac+, pea+s at /B-CI hours, returns to normal in three to 'our days. =Myocardial inMuries such as angina(chest pain) or pericarditis (in'lammation o' membrane around heart) do not increase .6% levels. 4ormal .6%: Males@ /CGBF units per liter (5(") or F.BHGF.HH micro+ats per liter (mc+at(")7 -emales@ /FGHK 5(" or F./LGF.KF mc+at(" High 0alue#: Nery high levels o' .6% may be caused by: ,ecent or severe liver damage, such as hepatitis caused by a viral in'ection or drug reaction7 3ecay o' a large tumor (necrosis)7 6hoc+. Moderately high levels o' .6% may be caused by: "ong-term (chronic) diseases that a''ect the liver, such as cirrhosis7 . heart attac+ or heart 'ailure7 .lcohol abuse7 !aving ta+en high doses o' vitamin .7 1idney or lung damage7 Mononucleosis7 3uchenne muscular dystrophy7 6ome types o' cancer7 . rare autoimmune disease that a''ects muscles (myositis). 6lightly high levels o' .6% may be caused by: -atty deposits in the liver7 .lcohol abuse7 .n overdose o' acetaminophen (%ylenol)7 Many medicines, such as statins, antibiotics, chemotherapy, aspirin, narcotics, and barbiturates. )$"OO3 D"5*O6# %#6%: measures the amount o' glucose (type o' sugar, main source o' energy) in blood. 2nsulin @ hormone that helps your bodys cells use glucose7 produced in pancreas and released into the blood when amount o' glucose in the blood. 4ormally, blood glucose levels a'ter you eat. ancreas releases insulin so that blood glucose levels are moderated. $lood glucose over time can damage eyes, +idneys, nerves and blood vessels. Fa#ting Blood Sugar&FBS' G measures blood glucose a'ter you have not eaten 'or at least I hours. 2t is o'ten the 'irst test done to chec+ 'or pre-diabetes and diabetes. 1 Hour 2o#t%randial Blood Sugarmeasures blood glucose two hours a'ter starting to eat a meal. Rando" Blood Sugar&RBS' G measures blood glucose regardless o' last meal. 6everal measurements may be ta+en throughout the day. 5se'ul because glucose levels in healthy people do not vary widely

throughout the day. $lood glucose levels that vary widely may mean a problem. %his test is also called a casual blood glucose test. Oral gluco#e tolerance te#t3 diagnose pre-diabetes and diabetes. 6eries o' blood glucose measurements ta+en a'ter drin+ing a sweet li;uid that contains glucose. 5sually only used to diagnose diabetes that occurs during pregnancy (gestational diabetes). $lood glucose tests are done to chec+ 'or diabetes, monitor treatment o' diabetes, chec+ gestational diabetes. 4ormal: LF to /FF mg(dl (J.K mmol(l) re-diabetes( 2mpaired Dlucose %olerance: /F/-/BK mg(dl 3iagnosis o' diabetes :more than /BK mg(dl 0 $"OO3 5,#. 42%,OD#4 ($54): measure amount o' nitrogen in the blood in the 'orm o' urea7 measurement o' renal 'unction. 5rea is a by- product 'rom metabolism o' proteins by the liver and is removed 'rom the blood by the +idneys. 5rea is made in the liver and passed out o' your body in the urine. . blood urea nitrogen ($54) test measures the amount o' urea in your blood. $54 test may be done with a blood creatinine test. $54 test is done to see i' your +idneys are wor+ing properly and to be able to maintain a healthy +idney. =1idney disease can @ $54 and creatinine levels. Increased BUN /. Medications .minoglycosides 3iuretics "ithium *orticosteroids B. Dastrointestinal $leeding H. 3ecreased ,enal blood 'low 6hoc+ *ongestive !eart 'ailure Myocardial 2n'aretion C. ,enal 3isease Dlomerulonephritis yelonephritis 3iabetic 4ephropathy J. 5rinary %ract Obstruction Decreased BUN /. "iver disease B. oor nutrition H. Overhydration C. %hird trimester o' pregnancy 4ormal $54: /F-BF mg(dl 'or adults7 J-/I mg(dl 'or children7 H-/B mg(dl 'or newborns, and B/-CF mg(dl 'or cord blood. . critical value o' /FF mg(dl indicates serious impairment o' renal 'unction. Creatinine: *reatinine and creatinine clearance tests measure level o' the waste

product creatinine in your blood and urine. %ell how well your +idneys are wor+ing. %he substance creatine is 'ormed when 'ood is changed into energy through a process called metabolism. *reatine is bro+en down into another substance called creatinine, which is ta+en out o' your blood by the +idneys and then passed out o' your body in urine. 6ee a picture o' the +idneys . *reatinine is made at a steady rate and is not a''ected by diet or by normal physical activities. 2' your +idneys are damaged and cannot wor+ normally, the amount o' creatinine in your urine goes down while its level in your blood goes up. %hree types o' tests on creatinine can be done: Blood creatinine le0el %he blood creatinine level shows how well your +idneys are wor+ing. . high creatinine level may mean your +idneys are not wor+ing properly. %he amount o' creatinine in the blood depends partly on the amount o' muscle tissue you have7 men generally have higher creatinine levels than women. Creatinine clearance te#t . creatinine clearance test measures how well creatinine is removed 'rom your blood by your +idneys. . creatinine clearance test gives better in'ormation than a blood creatinine test on how well your +idneys are wor+ing. . creatinine clearance test is done on both a blood sample and on a sample o' urine collected over BC hours (BC-hour urine sample). B$N3to3creatinine ratio &B$N:creatinine' $54:creatinine can help your doctor chec+ 'or problems (li+e dehydration) that may @ abnormal $54 and creatinine levels. . blood creatinine level or a creatinine clearance test is done to: 6ee i' your +idneys are wor+ing normally. 6ee i' your +idney disease is changing. 6ee how well the +idneys wor+ in people who ta+e medicines that can cause +idney damage. *hec+ 'or severe dehydration. 3ehydration generally causes $54 levels O creatinine levels @ !igh $54-tocreatinine ratio. 1idney disease or bloc+age o' the 'low o' urine 'rom your +idney causes both $54 and creatinine levels. 4ormal ,esults: Men- F.I to /.B mg(dl7 ?omen- F.K to F.E mg(dl $ric Acid in Blood %he blood uric acid test measures the amount o' uric acid in a blood sample. 5ric acid is produced 'rom the natural brea+down o' your body8s cells and 'rom the 'oods you eat.

Most o' the uric acid is 'iltered out by the +idneys and passes out o' the body in urine. . small amount passes out o' the body in stool. $ut i' too much uric acid is being produced or i' the +idneys are not able to remove it 'rom the blood normally, the level o' uric acid in the blood increases. !igh levels o' uric acid in the blood can cause solid crystals to 'orm within Moints. %his causes a pain'ul condition called gout. 2' gout remains untreated, these uric acid crystals can build up in the Moints and nearby tissues, 'orming hard lumpy deposits called tophi. !igh levels o' uric acid may also cause +idney stones or+idney 'ailure. . uric acid blood test is done to: !elp diagnose gout. *hec+ i' uric acid levels cause +idney stones. *hec+ i' medicine thaturic acid is wor+ing. *hec+ uric acid levels in people who are undergoing chemotherapy or radiation therapy. %hese treatments destroy cancer cells that then may lea+ uric acid into the blood. (4ormal Nalues 'all between H.F and L.F mg(dl) Chole#terol Te#t#: *holesterol: provide stability to the outer membranes o' our bodies8 cells. $ut cholesterol can have also harm'ul e''ects. "3" PbadP cholesterol can deposit in blood vessel walls. Over years, "3" cholesterol and other substances clog arteries in the process called atherosclerosis. .rteries in the heart narrowed by atherosclerosis can then develop sudden blood clots, causing heart attac+s. !aving high total cholesterol, high bad cholesterol, or low good cholesterol places you at higher than average ris+ 'or heart disease. 1nowing about high cholesterol is the 'irst step toward lowering it, which reduces your ris+. %hat8s the reason to get a cholesterol test. O0er0ie4 o, Chole#terol Te#t# %here are multiple 'orms o' cholesterol circulating in the blood. Narious 'orms o' cholesterol A other 'ats in the blood @ lipids. 3octors measure and diagnose cholesterol with a simple blood test, o'ten called a lipid pro'ile a'ter 'asting 'or E-/B hours (eliminate contribution o' 'at recently eaten. %ests 'or cholesterol provide results 'or 'our di''erent types o' lipids. %otal cholesterol "3" (low-density lipoprotein), the Pbad cholesterolP !3" (high-density lipoprotein), the Pgood cholesterolP %riglycerides, another 'orm o' 'at in the blood

6ome lipid panels provide more detailed in'ormation, with in'ormation on the presence and si)es o' various 'at particles in the blood. ,esearch is ongoing into the possible contribution to heart disease o' these 'actors. *urrently, there is no consensus on when more advanced testing o' 'at particle si)e is necessary. -or total cholesterol: BFF milligrams per deciliter (mg(d") or less is considered normal (good). BF/ to BCF mg(d" is borderline. Dreater than BCF mg(d" is high. =-or !3" (Pgood cholesterolP), better: !3" KF mg(d" or higher is good -- it protects against heart disease. !3" between CF and JE mg(d" are acceptable. "ess than CF mg(d" !3" is low, increasing the ris+ o' heart disease. -or "3" (Pbad cholesterolP), better: "3" Q /FF mg(d" is optimal "3" o' /FF to /BE mg(d" or @ nearoptimal. "3" between /HF and /JE @ borderline high. "3" cholesterol o' /KF mg(d" or more @ high. "3" o' /EF mg(d" or more @ very high. 0 "iver -unction %ests ("-%s) *linical biochemical laboratory blood assaysin'o about state o' patients liver. 3one by M% on patients serum( plasma ('rom phlebotomy.) *an be used to: (/) detect liver disease, (B) distinguish among di''erent types o' liver disorders, (H) gauge the e<tent o' liver damage and (C) 'ollow the response to treatment. .lbumin (.lb): rotein made only by liver. @chronic liver disease. .lanine %ransaminase (."%): 6erum Dlutamic yruvate %ransaminase (6D %) or .lanine .minotrans'erase (.".%). #n)yme present in hepatocytes (liver cells). @ liver damage, (."% lea+ed into the blood) .spartate %ransminase (.6%): 6erum Dlutamic O<aloacetic %ransaminase (6DO%) or .spartate .minotrans'erase (.6.%). @acute liver damage, but is present in ,$*s and cardiac and s+eletal muscle (R .6%@not speci'ic to the liver) =,atio o' .6% to ."% is sometimes use'ul in di''erentiating between causes o' liver damage .l+aline hotophatase (." ): #n)yme in the cells lining the biliary ducts o' the liver. ." in plasma @ certain liver diseases. ." in bone and placental tissue, R in growing children and elderly patients.

%otal $ilirubin (%$2"): $rea+down product o' heme (part o' hemoglobin). "iver clears blood o' bilirubin. *an signal the (/)prehepatic (B) hepatic (H) posthepatic problems 3irect $ilirubin (conMugated bilirubin): 3iagnosis is narrowed down 'urther by loo+ing at the levels o' direct bilirubin. =2' direct (i.e. conMugated) bilirubin is normal, then problem is an e<cess o' unconMugated bilirubin, and the location o' the problem is upstream o' bilirubin e<cretion. 6uspect: !emolysis, viral hepatitis, or cirrhosis. =2' direct bilirubin, liver is conMugating bilirubin normally, but cant e<crete it. 6uspect: $ile duct obstruction by gallstones or cancer. Damma Dlutamyl %ranspeptidase (DD%): even in minor, sub-clinical levels o' liver dys'unction. !elp'ul in identi'ying the cause o' an isolated elevation in ." . (DD% is raised in chronic alcohol to<icity). 2re%aration o, the 2atient N#42 54*%5,# ,O*#35,# %he venipuncture procedure is comple<, re;uiring both +nowledge and s+ill to per'orm. #ach phlebotomist generally establishes a routine that is com'ortable 'or her or him. 6everal essential steps are re;uired 'or every success'ul collection procedure: /. 2denti'y the patient. B. .ssess the patient8s physical disposition (i.e. diet, e<ercise, stress, basal state). H. *hec+ the re;uisition 'orm 'or re;uested tests, patient in'ormation, and any special re;uirements. C. 6elect a suitable site 'or venipuncture. J. repare the e;uipment, patient > puncture site. K. er'orm the venipuncture. L. *ollect sample in appropriate container. I. ,ecogni)e complications associated with the phlebotomy procedure. E. .ssess the need 'or sample recollection and(or reMection. /F. "abel collection tubes at bedside( drawing area. //. romptly send specimens with re;uisition to lab. #;uipment needed: #vacuated *ollection %ubes - %he tubes are designed to 'ill with a predetermined volume o' blood by vacuum. %he rubber stoppers are color coded according to the additive that the tube contains. Narious si)es are available. $lood should NE5ER be poured 'rom one tube to another since the tubes can have di''erent additives or coatings 4eedles - %he gauge number indicates the bore si)e: the larger the gauge number, the smaller the needle bore. 4eedles are available 'or evacuated systems and 'or

use with a syringe, single draw or butter'ly system. !older(.dapter - use with the evacuated collection system. %ourni;uet - ?ipe o'' with alcohol and replace 're;uently. .lcohol ?ipes - LFS isopropyl alcohol. ovidone-iodine wipes(swabs - 5sed i' blood culture is to be drawn. Dau)e sponges - 'or application on the site 'rom which the needle is withdrawn. .dhesive bandages ( tape - protects the venipuncture site a'ter collection. 4eedle disposal unit - needles should 4#N#, be bro+en, bent, or recapped. 4eedles should be placed in a proper disposal unit 2MM#32.%#"& a'ter their use. Dloves - can be made o' late<, rubber, vinyl, etc.7 worn to protect the patient and the phlebotomist. 6yringes - may be used in place o' the evacuated collection tube 'or special circumstances. 2rocedural i##ue# .%2#4% ,#".%2O46 .43 23#4%2-2*.%2O4: %he phlebotomist8s role @ pro'essional, courteous, and understanding. Dreet patient, identi'y yoursel' and indicate the procedure that will ta+e place. #''ective verbal and nonverbal communication. roper patient identi'ication M.43.%O,&. 2' an in-patient is able to respond, as+ 'or a 'ull name and always chec+ the armband or bracelet 'or con'irmation. -O NOT -RA6 BLOO- IF THE ARMBAN- OR BRACELET IS MISSIN7! -or an in-patient, nursing sta'' can be contacted to aid in identi'ication prior to proceeding. .n out-patient must provide identi'ication other than the verbal statement o' a name. 5sing the re;uisition 'or re'erence, as+ a patient to provide additional in'ormation such as a surname or birthdate. . government issued photo i.d. card such as a driver8s license can aid in resolving identi'ication issues. 2' possible, spea+ with the patient during the process. .t easeless 'ocused on the procedure. .lways than+ the patient and e<cuse yoursel' courteously when 'inished. -ia(ete# Mellitu# O'ten simply re'erred to as dia(ete#, it is a group o' metabolic diseases in which a person has blood sugar, either: body does not produce enough insulin, or because cells do not respond to the insulin that is produced. %his high blood sugar produces the classical symptoms: polyuria ('re;uent urination), polydipsia (increased thirst) and polyphagia (increased hunger).

Ty%e#: Ty%e dia(ete#: body 'ails to produce insulin7 re;uires the person to inMect insulin. (2nsulin-dependent diabetes mellitus, 233M, and Muvenile diabetes.) Ty%e 1 dia(ete#: insulin resistance7 cells 'ail to use insulin properly, sometimes combined with an absolute insulin de'iciency. (4on-insulindependent diabetes mellitus, 4233M, adult-onset diabetes mellitus. .O3M.) 7e#tational dia(ete#: pregnant women, never had diabetes, have blood glucose level. May precede development o' type B 3M. Congenital dia(ete#: genetic de'ects o' insulin secretion Steroid dia(ete#: induced by high doses o' glucocorticoids Sy"%to"#: = 6ymptoms may develop rapidly (wee+s or months) in type / diabetes while in type B diabetes they usually develop much more slowly and may be subtle or absent. rolonged blood glucoseglucose absorptionshape o' the lenses o' the eyes changeresulting in vision changes7 sustained sensible glucose control usually returns the lens to original shape. $lurred vision is a common complaint leading to a 3M diagnosis7 type / always suspected in cases o' rapid vision change, type B changes are generally more gradual, but should still be suspected. 6ome untreated 3M patients 'eel: 'atigue, nausea > vomiting. . rarer but e;ually severe possibility is hyperosmolar non+etotic state, more common in type B 3M and is mainly the result o' dehydration (patient drin+ing large amounts o' sugary drin+s.) . number o' s+in rashes can occur in diabetes that are collectively +nown as diabetic dermadromes. ,elative(absolute insulin de'iciency @ weight despite appetite. Cau#e#: %ype / diabetes: pancreas undergoes an autoimmune attac+ by the body itsel' > rendered incapable o' ma+ing insulin. %he patient with type / diabetes must rely on insulin medication 'or survival. .utoimmune are mista+enly manu'actured by the immune system which are directed to the pancreas and damages the beta cells that are responsible 'or insulin production. %his type o' condition is believed to be genetically inherited. #<posure to certain viral in'ections (mumps and *o<sac+ie viruses) or other environmental to<ins may serve to trigger

abnormal antibody responses that damage the pancreas cells where insulin is made. 6ome o' the antibodies seen in type / 3M include anti-islet cell antibodies, antiinsulin antibodies and anti-glutamic decarbo<ylase antibodies. %hese antibodies can be measured in the maMority o' patients, and may help determine which individuals are at ris+ 'or developing type / 3M. %ype B diabetes 2n type B diabetes, patients can still produce insulin, but do so relatively inade;uately 'or their body8s needs, particularly in the 'ace o' insulin resistance as discussed above. 2n many cases this actually means the pancreas produces larger than normal ;uantities o' insulin. . maMor 'eature o' type B diabetes is a lac+ o' sensitivity to insulin by the cells o' the body (particularly 'at and muscle cells). %he liver in these patients continues to produce glucose through a process called gluconeogenesis despite elevated glucose levels. %he control o' gluconeogenesis becomes compromised. Most o' these cases are a direct result o' poor eating habits, higher body weight, and lac+ o' e<ercise. ?hile there is a strong genetic component to developing this 'orm o' diabetes, there are other ris+ 'actors - the most signi'icant o' which is obesity. 3iabetes can occur temporarily during pregnancy. 6igni'icant hormonal changes during pregnancy can lead to blood sugar elevation in genetically predisposed individuals. $lood sugar elevation during pregnancy is called gestational diabetes. P6econdaryP diabetes re'ers to elevated blood sugar levels 'rom another medical condition. 6econdary diabetes may develop when the pancreatic tissue responsible 'or the production o' insulin is destroyed by disease, such as chronic pancreatitis (in'lammation o' the pancreas by to<ins li+e e<cessive alcohol), trauma, or surgical removal o' the pancreas. 3iabetes can also result 'rom other hormonal disturbances, such as e<cessive growth hormone production 2n acromegaly, a pituitary gland tumor at the base o' the brain causes growth hormone, leading to hyperglycemia. 2n *ushing8s syndrome, the adrenal glands produce an cortisol, which promotes blood sugar. -iagno#i#: The ,a#ting (lood gluco#e &#ugar' te#t@pre'erred way to diagnose. #asy to per'orm and convenient. .'ter the person

has 'asted overnight (T I hours), a single sample o' blood is drawn and sent to the laboratory 'or analysis. %his can also be done accurately in a doctor8s o''ice using a glucose meter. =4ormal 'asting plasma glucose levels are Q/FF milligrams per deciliter(mg(dl). =-asting plasma glucose levels o' /BK mg(dl on twoA tests on di''erent days@diabetes. =. random blood glucose test can also be used to diagnose diabetes. . blood glucose level o' BFF mg(dl or higher indicates diabetes. Oral gluco#e tolerance te#t &O7TT' (non-routine) is a gold standard 'or ma+ing the diagnosis o' type B diabetes. ?ith an oral glucose tolerance test, the person 'asts overnight (T I but Q/K hours). -asting plasma glucose is then tested. .'ter this test, the person receives LJ grams o' glucose (/FF grams 'or pregnant women). %here are several methods employed by obstetricians to do this test, but the one described here is standard. 5sually, the glucose is in a sweettasting li;uid that the person drin+s. $lood samples are ta+en at speci'ic intervals to measure the blood glucose. -or the test to give reliable results: person must be in good health (not ill7 not even a cold). person should be normally active (not lying down as an inpatient in a hospital) person should not be ta+ing medicines that could a''ect the blood glucose. -or three days be'ore the test, the person should have eaten a diet high in carbohydrates (BFF-HFF grams per day). %he morning o' the test, the person should not smo+e or drin+ co''ee. Medication# Met,or"in Denerally recommended as a 'irst line treatment 'or type B 3M as there is good evidence that it mortality. ,outine use o' aspirin not 'ound to improve outcomes in uncomplicated diabetes. In#ulin %ype / 3M is typically treated with combinations o' regular and 4 ! insulin, or synthetic insulin analogs. ?hen insulin is used in type B diabetes, a long-acting 'ormulation is usually added initially, while continuing oral medication 3oses o' insulin are then increased to e''ect. *hemist- blood chemistry - technical 1! He"atology: =$lood with anti-coagulant: pac+ed red cellA ".6M. =$lood without anti-coagulant: solid blood clotA6#,5M (6erum: no 'ibrinogen. roteins, electrolytes, antibodies, antigens, hormones.) 0*OM "#%# $"OO3 *O54% (*$*)

,$*, ?$*, platelets, hemoglobin, hematocrit ?$*@viral in'ection, i' chemotherapy ?$*@bacterial in'ection *an be done through: ric+ Method Nenipuncture .nte-cubital 'ossae o' the 'orearm Midcubital *ephalic $asilic %he complete blood count or *$* test is used as a broad screening test to chec+ 'or such disorders as anemia, in'ection, and many other diseases. 2t is actually a panel o' tests that e<amines di''erent parts o' the blood and includes the 'ollowing: ?hite blood cell (?$*) count: actual number o' white blood cells per volume o' blood. ?hite blood cell di''erential: types o' white blood cells present. %here are 'ive di''erent types o' white blood cells, each with its own 'unction in protecting us 'rom in'ection: lymphocytes, monocytes, neutrophils (.1.: segs, M4s, granulocytes, grans), eosinophils, and basophils. ,ed blood cell (,$*) count : actual number o' red blood cells per volume o' blood. !emoglobin: amount o' OB-carrying protein in the blood. !ematocrit: percentage o' red blood cells in a given volume o' whole blood. latelet count: number o' platelets in a given volume o' blood. Mean platelet volume (M N): machinecalculated measurement o' the average si)e o' your platelets. Mean corpuscular volume (M*N): average si)e o' your ,$*s. Mean corpuscular hemoglobin (M*!): average amount o' OB-carrying hemoglobin inside a red blood cell. Mean corpuscular hemoglobin concentration (M*!*): average concentration o' hemoglobin inside a red cell. ,ed cell distribution width (,3?): variation in the si)e o' ,$*s. ,eticulocyte count: how 'ast red blood cells called reticulocytes are made by the bone marrow and released into the blood. ,eticulocytes are in the blood 'or about B days be'ore developing into mature red blood cells. 4ormal@ 9/-BS o' the ,$*s in the blood are reticulocytes. 0*oagulation studies (clotting 'actors) 5sed by health care providers to determine i':

level o' 'actor@low(absentreduced clot 'ormation and bleeding level o' 'actor@too hightoo much clot 'ormation > thrombosis. *oagulation 'actors measured to monitor levels during therapy. %ests are used as screening tools to determine whether one has a coagulation problem. *oagulation 'actor tests may be ordered when: #<cessive bleeding or bruising is e<perienced. rolonged % or %%. 2nherited 'actor de'iciency is suspected. ($leeding episodes begin early in li'e or when a close relative has an inherited 'actor de'iciency. =2' inherited de'iciency is suspected, test other 'amily members to help con'irm diagnosis and establish whether they may be carriers or have the de'iciency (asymptomatic(less severe).) .c;uired condition that causes bleeding, (e<: vitamin 1 or liver disease) is suspected. . patient with a +nown de'iciency (monitor 'actor de'iciency > evaluate treatment e''ectivity. 4ormal coagulation 'actor activity9 normal clotting 'unction. "ow activity o' one or more coagulation 'actors 9impaired clotting ability. #ach coagulation 'actor needs a certain ;uantity 'or normal clotting, but the level re;uired is di''erent 'or each 'actor. ,esults are 're;uently reported as a percentage with /FFS being normal. #<ample, a 'actor N222 that is HFS is abnormally low. 3e'iciencies in coagulation 'actors may be ac;uired or inherited, mild or severe, permanent or temporary. %hose that are inherited are rare and tend to involve only one 'actor, which may be reduced or absent. !emophilia . and $ @ most common e<amples o' inherited disorders. U-lin+ed de'iciencies o' 'actors N222 and 2U that occur almost e<clusively in men (women @ asymptomatic carriers who may have mild bleeding). Other inherited 'actor de'iciencies, no U chromosome involved, are 'ound e;ually in both se<es. 6everity o' symptoms e<perienced depends on the 'actor involved and amount available. 6ymptoms may vary 'rom episode to episode, 'rom e<cessive bleeding a'ter dental procedures to severe recurrent bleeding into Moints or muscles. atients with a modest reduction in coagulation 'actor level may e<perience 'ew symptoms and may discover their de'iciency as an adult a'ter a surgical procedure or trauma or

during screening that includes a rothrombin %ime ( %) or artial %hromboplastin %ime ( %%) test. %hose with severe 'actor de'iciencies may have their 'irst bleeding episode very early7 'or e<ample, a male in'ant with a de'iciency o' -actor N222, 2U, or U222 may bleed e<cessively a'ter circumcision. .c;uired de'iciencies may be due to chronic diseases, such as liver disease or cancer7 to an acute condition such asdisseminated intravascular coagulation (32*), which uses up clotting 'actors at a rapid rate7 or to a de'iciency in vitamin 1 or treatment with a vitamin 1 antagonist li+e war'arin (the production o' 'actors 22, N22, 2U, and U re;uire vitamin 1). 2' more than one clotting 'actor is decreased, it is usually due to an ac;uired condition. -actors may be decreased because o': 0 consumption due to e<tensive clotting 0 liver disease 0 uremia (abnormally high level o' nitrogentype wastes in the blood) 0 some cancers 0 bone marrow disorders 0 e<posure to sna+e venom 0 vitamin 1 de'iciency 0 anticoagulation therapy 0 accidental ingestion o' the anticoagulant war'arin 0 multiple blood trans'usions (stored units o' blood lose some o' their clotting 'actors) #levated levels o' several 'actors are seen in situations o' acute illness, stress, or in'lammation. 6ome patients have persistent elevations o' 'actor N222 that may be associated with an increased ris+ o' venous thrombosis. Te#t# o, the 5a#cular 2latelet 2ha#e o, He"o#ta#i# 0$"##324D %2M#: 6mall slit is made in the s+in, the hemostatic mechanisms necessary 'or coagulation are activated. 4o pressure, bleeding usually stops within L-E minutes. 5se disposable template that produces a uni'orm incision. %he incision, either hori)ontal or vertical, is placed on the lateral aspect o' the 'orearm, about J cm below the antecubital 'ossa, a'ter a blood pressure cu'' has been in'lated to appro<imately CF mm !g. $lood may be absorbed o'' the s+in, but .NO23 ,#665,#. %he time is measured 'rom the moment o' incision to the moment bleeding stops. %ime varies: commercial template used, incision direction, location on arm. 3u+e Method G on the earlobe or 'ingertips 2vy Method G on the 'orearm %he vascular platelet phase o' hemostasis @ primary vasoconstriction that serves to

blood 'low A adherence o' platelets to the ruptured endothelium (adhesion) and each other (aggregation). %his platelet aggregate, called the platelet plug, stops the bleeding and 'orms a matri< 'or the clot. %he bleeding time is an e<cellent screening test 'or the vascular platelet phase o' hemostasis. 2t depends on an intact vasospastic response in a small vessel and an ade;uate number o' 'unctionally active platelets. *linical 6igni'icance: atients with abnormalities o' the vascular platelet phase o' hemostasis present with purpura (petechiae and ecchymoses) and spontaneous bruising. %hey may have mucosal bleeding and 'undus hemorrhages. *ommonly, the problem is either thrombocytopenia, easily evaluated by a platelet count, or abnormal platelet 'unction, which can be diagnosed with platelet 'unction studies. %he most common ac;uired platelet 'unction abnormalities are drug induced (aspirin and the nonsteroidal anti-in'lammatory agents) and uremia. %he most common hereditary abnormality is von ?illebrand8s disease. Te#t# o, the Coagulation Ca#cade %hese in vitro teststhe activated partial thromboplastin time (a %%), prothrombin time ( %), and thrombin time (%%)measure the time elapsed 'rom activation o' the coagulation cascade at di''erent points to the generation o' 'ibrin. 0.*%2N.%#3 .,%2." %!,OM$O ".6%24 %2M#: time necessary to generate 'ibrin 'rom initiation o' the intrinsic pathway. .ctivation o' 'actor U22 is accomplished with an e<ternal agent (e.g., +aolin) capable o' activating 'actor U22 without activating 'actor N22. 6ince platelet 'actors are necessary 'or the cascade to 'unction normally, the test is per'ormed in the presence o' a phospholipid emulsion that ta+es the place o' these 'actors. %he classic partial thromboplastin time depends on contact with a glass tube 'or activation. 6ince this is considered a di''icult variable to control, the PactivatedP test uses an e<ternal source o' activation. %echni;ue: *itrated plasma, an activating agent, and phospholipid are added together and incubated at HLV*. *alcium is added, and the time necessary 'or the clumping o' +aolin is measured. %he normal time is usually reported as less than HF to HJ seconds depending on the techni;ue used. 2n 'act, there is a normal range o' about /F seconds (e.g., BJ to HJ), and decreased values (PshortP) may also be abnormal. $asic 6cience: %his test is abnormal in the presence o' reduced ;uantities o' 'actors U22, 2U, U2, N222, U, N, prothrombin, and 'ibrinogen (all integral parts o' the PintrinsicP and

PcommonP pathway. 2t is usually prolonged i' a patient has less than appro<imately HFS normal activity. 2t can also be abnormal in the presence o' a circulating inhibitor to any o' the intrinsic pathway 'actors. %he di''erentiation o' inhibitors 'rom 'actor depletion is important and can best be accomplished by a mi<ing study in which patient and normal plasma are combined in a /:/ ratio and the test is repeated on the mi<ed sample. 2' the abnormal value is corrected completely, the problem is 'actor de'iciency. 2' the result does not change or the abnormality is corrected only partially, an inhibitor is present. %his di''erence stems 'rom the above mentioned 'act that the a %% will be normal in the presence o' JFS normal activity. *linical 6igni'icance: %he a %% is a good screening test 'or inherited or ac;uired 'actor de'iciencies. 2nherited disorders including classic hemophilia . ('actor N222 de'iciency) and hemophilia $ ('actor 2U de'iciency, or *hristmas disease) are well+nown diseases in which the a %% is prolonged. Other intrinsic and common pathway 'actors may also be congenitally absent. %hese conditions are rare but have been described 'or all 'actors. . number o' +indreds with abnormalities o' 'actor U22 activation have been described. %hey are usually associated with a prolonged a %% without clinical signs o' bleeding. .c;uired 'actor de'iciency is common. Nitamin 1 de'iciency, liver dys'unction, and iatrogenic anticoagulation with war'arin are most common. -actor depletion may also occur in the setting o' disseminated intravascular coagulation (32*), prolonged bleeding, and massive trans'usion. . prolonged a %% that cannot be completely normali)ed with the addition o' normal plasma can be e<plained only by the presence o' a circulating inhibitor o' coagulation. %he presence o' these inhibitors is almost always ac;uired, and their e<act nature is not always apparent. -rom a clinical point o' view, the most common inhibitors should be considered antithrombins. %hese compounds inhibit the activity o' thrombin on the conversion o' 'ibrinogen to 'ibrin. %he two most common inhibitors are heparin, which acts through the naturally occurring protein antithrombin 222 (.% 222), and 'ibrin degradation products (-3 ), 'ormed by the action o' plasmin on the 'ibrin clot and usually present in elevated concentrations in 32* and primary 'ibrinolysis. Other inhibitors appear to be antibodies. %he easiest to understand is the antibody against 'actor N222 in patients with hemophilia . treated with 'actor N222 concentrate. 2nhibitors against other 'actors have been

described with a variety o' diseases that 'ollow a variable course. ?hen characteri)ed, they have been immunoglobulins. . particular problem may be seen in patients su''ering 'rom systemic lupus erythematosus. %hese patients may present with a prolonged a %% without evidence o' bleeding. 6ome present with thrombosis. %he abnormality cannot be corrected with normal plasma and has been re'erred to as the Plupus anticoagulant.P %his phenomenon does not represent an in vivo problem with the coagulation cascade. ,ather, it is a laboratory abnormality caused by the presence o' a serum constituent that inter'eres with the in vitro partial thromboplastin test. Occasionally the reported value o' the a %% will be lower than normal. %his PshortenedP time may re'lect the presence o' increased levels o' activated 'actors in conte<t o' a Phypercoagulable state.P 2t is seen in some patients in the early stages o' 32* but should not be considered diagnostic 'or that entity. 0 ,O%!,OM$24 %2M#: time necessary to generate 'ibrin a'ter activation o' 'actor N22. 2t measures the integrity o' the Pe<trinsicP and PcommonP pathways ('actors N22, N, U, prothrombin, and 'ibrinogen). %echni;ue: *itrated plasma and an activating agent (usually thromboplastin e<tracted 'rom animal brain) are incubated at HLV*. %he plasma is recalci'ied and the time is measured until 'ibrin 'ilaments are observed. #ach laboratory has its own normal value, usually between /B and /J seconds. $asic 6cience: .s with the interpretation o' a prolonged a %%, a prolonged % may re'lect either 'actor de'iciency or a circulating inhibitor o' coagulation. %he distinction is made by repeating the test a'ter a /:/ mi< with normal plasma. %est is more sensitive than the a %% 'or de'icient levels o' 'actors, and a relatively small drop in 'actor N22 levels may prolong the %. *linical 6igni'icance: 2nherited de'iciency o' 'actor N22 is a rare bleeding disorder characteri)ed by a prolonged % and a normal a %%. %he % completely corrects when mi<ed with normal plasma. .c;uired de'iciencies usually related to liver disease, war'arin therapy, or depletion secondary to consumptive coagulopathy, severe bleeding, or massive trans'usion. *irculating inhibitors most o'ten directed at 'actor U or thrombin. Most common are heparin or products o' 'ibrinolysis. 2n their presence the prolonged % cannot be completely corrected to normal in a /:/ mi<ing study. 0%!,OM$24 %2M#: %his test measures the time necessary to drive the reaction o' 'ibrinogen to 'ibrin in the presence o'

thrombin. 2t measures the integrity o' this reaction and isolates an abnormality to either a decrease in normal 'ibrinogen or an inhibitor to its activation. %echni;ue: *itrated plasma is incubated at HLV* and thrombin is added to the solution. %ime is measured 'rom the addition o' thrombin to the generation o' 'ibrin 'ilaments. *alcium is unnecessary. $asic 6cience: .bnormalities can be e<plained in one o' three ways: de'icient 'ibrinogen (Q /FF mg(dl), abnormal 'ibrinogen, or an inhibitor to the reaction. .s with other tests o' the coagulation cascade, i' a /:/ mi<ing study normali)es the prolonged time, one is dealing with 'actor de'iciency. .s it pertains to 'ibrinogen, however, one must distinguish a decrease in normal 'ibrinogen 'rom the production o' an abnormal 'ibrinogen (dys'ibrinogenemia). *linical 6igni'icance: .c;uired de'iciency o' 'ibrinogen is usually due to a consumptive coagulopathy or, less o'ten, severe liver disease. !ereditary de'iciencies e<ist, but with variable clinical presentations. .'ibrinogenemia is an o'ten 'atal childhood condition. .bnormal 'ibrinogen (dys'ibrinogenemia) can be ac;uired or inherited. %he ac;uired 'orm is usually 'ound in association with severe liver disease, but has been reported in other diseases. %he congenital 'orm is rare, usually autosomal dominant. . discordance between immunologic and physiologic measurements o' 'ibrinogen is the +ey to diagnosis. %he most common ac;uired inhibitors o' this reaction are heparin and 'ibrin degradation products (-3 ). %he e''ect o' heparin can be eliminated by cataly)ing the reaction with reptilase, which, unli+e thrombin, is insensitive to heparin. -3 are commonly seen in consumptive coagulopathies and primary 'ibrinolytic states. Te#t# o, Fi(rinoly#i# and the Mechani#"# That Control He"o#ta#i# %he coagulation cascade is an e''icient system 'or generating 'ibrin. #;ually important is the system that limits this process, neutrali)es activated 'actors, stops propagation o' the clot, and +eeps the process o' coagulation con'ined to the area o' endothelial rupture. 0-2$,24O"&626: accomplished by the action o' plasmin on 'ibrin polymer. lasmin is generated 'rom plasminogen (produced by the liver) by the action o' plasminogen activators. %hese compounds are present in endothelial cells, and the reaction is accelerated in the presence o' 'ibrin. %hus the generation o' 'ibrin appears to turn on the 'ibrinolytic process and locali)e the 'ormation o' the 'ibrin gel. 0-2$,24 3#D,.3.%2O4 ,O35*%6: .s a mar+er o' 'ibrinolysis, -3 , (also +nown as

'ibrin split products (-6 ),) can be ;uanti'ied by a test based on late< agglutination. %he test uses antibodies to -6 which are measured using serial dilutions. %echni;ue: 6erum is prepared in a series o' dilutions (e.g., :, W, /(/K). "ate< particles onto which have been absorbed antibodies to -6 are added. 2' agglutination is seen, the test is positive at that dilution. %he test is reported as the most dilute sample that agglutinates. %he normal value is less than W to X. $asic 6cience: .lthough well standardi)ed and easy to per'orm, the -6 value may be di''icult to interpret. %he action o' thrombin on 'ibrinogen is to cleave the protein and produce smaller compounds called 'ibrinopeptides, plus the 'ibrin monomer. %his monomer polymeri)es to 'orm the 'ibrin gel. %he gel is stabili)ed by the action o' 'actor U222, activated by thrombin. %he action o' plasmin is to cleave both 'ibrinogen and 'ibrin. 2ts action is locali)ed by its activation at the site o' endothelial rupture, and the tight association o' plasminogen and 'ibrin. %he activity on 'ibrinogen 'orms small 'ragments, 3 and #. %he action on 'ibrin polymer is to 'orm larger 'ragments. %hese 'ragments are anticoagulants 'ormed at the site o' coagulation and serve to inhibit the action o' thrombin on 'ibrinogen to 'orm 'ibrin. $oth are also measured by the techni;ue described above. *linical 6igni'icance: 2ncreased -3 is the laboratory e<pression o' increased 'ibrinolysis. %his may be part o' a local problem o' 'ibrin generation such as brain trauma, chronic bleeding, vascular thrombosis, prostate surgery, uterine disorders, or malignancy, or a systemic process, usually 32*. atients with severe liver disease can have increased 'ibrinolysis on the basis o' poor clearance o' circulating plasminogen activators. Other *ontrol Mechanisms .t least two other systems are important in controlling the hemostatic process. .ntithrombin 222 (.% 222) is a protein produced by the liver. 2t binds thrombin and activated 'actor U irreversibly. %his interaction is accelerated by heparin, which may be associated with ruptured endothelial cells. .% 222 can be ;uanti'ied, and is congenitally absent in some patients prone to thrombosis. . similar protein (protein *) appears to inactivate 'actors N and N222 in a reaction cataly)ed by thrombin, locali)ed to the area o' endothelial rupture. 2t is a vitamin 1 dependent protein produced by the liver. 2ts measurement is still considered a research tool. $oth o' these proteins are locally activated

at the site o' endothelial rupture. %hey serve to prevent the escape into the circulation o' activated 'actors and limit propagation o' the clot. 0#,&%!,O*&%# 6#32M#4%.%2O4 ,.%#(#6,) %he #6, is a simple non-speci'ic screening test that indirectly measures the presence o' in'lammation in the body. 2t re'lects the tendency o' red blood cells to settle more rapidly in the 'ace o' some disease states, usually because o' increases in plasma 'ibrinogen, immunoglobulins, and other acute-phase reaction proteins. *hanges in red cell shape or numbers may also a''ect the #6,. Myelogenous leu+emia Y bone marrow, myeloblasts may appear in the blood. 6taining Methods Romanowsky stainingprototypical staining techni;ue: 'orerunner o' several distinct but similar methods, (Diemsa, Zenner, ?right, -ield, and "eishman stains) used to di''erentiate cells in pathologic specimens. May-Grunwald-Giemsa method - 5sed to study the adherence o' pathogenic bacteria to human cells. Leishman's method - 6uitable when stained blood is re;uired urgently or the routine stain is unavailable Field's stain method - 5sed 'or staining thin blood 'ilms in order to discover malarial parasites. Wright-Giemsa Stain Method - used in cytogenetics and 'or the histopathological diagnosis o' malaria and other parasites !ematologist- interprets slides, presence o' abnormal cells. (M% preps slides.) 8! Hi#to%athology: 6tudy o' the di''erent pathologic cells in the di''erent organs o' the bodies ($iopsy, autopsy, rush-'ro)en section Qimmediate e<amO) ,e'ers to the microscopic e<amination o' tissue to study the mani'estations o' disease. !istopathological e<amination o' tissues starts with #urgery9 (io%#y9 or auto%#y! %he tissue is removed 'rom body or plant and then placed in a 'i<ative which stabili)es the tissues to prevent decay. Most common 'i<ative @ 'ormalin (/FS 'ormaldehyde) Auto%#y: medical e<am o' the body o' a person who has died7 post-mortem e<aminationdetermines cause and manner o' death and evaluate any disease or inMury related to the death o' the patient(victim. .utopsies are per'ormed 'or either legal or medical purposes. .) "egal: 'orensic autopsy, victim is thoroughly e<amined to determine cause o' death, which may be relevant and help'ul in solving a murder case, or deducing that the victim simply died o' an accident.

$) Medical: the victim died 'rom a previously un+nown disease, (or a rare disease7) may help researchers determine cause o' disease via e<amining the condition o' the body. %ypes o' #<aminations 2nvolved: a) #<ternal #<amination: usually provides su''icient evidence to conclude the autopsy $ody is photographed 4ote +ind o' clothes and their position on the body #vidence such as residue, 'la+es o' paint or other material is collected 'rom e<ternal sur'aces o' the body G usually with ultraviolet light 6amples o' hair, nails, etc. and the body may also be radiographically imaged .ny wounds present are e<amined $ody is then cleaned, weighed, and measured in preparation 'or the internal e<amination . general description o' the body as regards ethnicity, se<, age, hair color and length, eye color and other distinguishing 'eatures (birthmar+s, old scar tissue, moles, etc.) is made - handheld voice recorder is usually used b) 2nternal #<amination: complete internal e<amination includes removal o' and dissection o' the chest, abdominal, and pelvic organs and the brain. %he e<amination o' the trun+ re;uires an incision 'rom the chest to the abdomen. %he removal o' the brain re;uires an incision over the top o' the head. Organs may be placed in 'ormalin 'or days to wee+s prior to dissection. %his is particularly important in the e<amination o' the brain 'or certain types o' diseases or inMuries. %issue samples are ta+en 'rom some or all o' the organs 'or e<amination under a microscope. 6amples o' blood, organs, and body 'luids may be removed and preserved to test 'or drugs or in'ection or to evaluate chemical composition or genetics. 6amples may include blood 'rom the heart or blood vessels, vitreous gel 'rom the eyes, bile 'rom the gallbladder, contents o' the stomach, urine, and tissues 'rom organs, such as the liver. 3etailed rocedure: i. "arge, deep, & Gshaped incision that is made 'rom shoulder to shoulder meeting at the breast bone and e<tends all the way down to the pubic bone. 2' 'emale, the &-incision is curved around bottom o' the breasts be'ore meeting the breast bone. ii. %he ne<t step is to peel bac+ the s+in, muscle and so't tissue using a scalpel. Once this is done, the chest 'lap is pulled up over the 'ace, e<posing the ribcage and nec+ muscles.

iii. %wo cuts are made on each side o' the ribcage, and then the ribcage is pulled 'rom the s+eleton a'ter dissecting the tissue behind it with a scalpel iv. ?ith the organs e<posed, a series o' cuts are made that detach the laryn<, esophagus, various arteries and ligaments. 4e<t, the medical e<aminer severs the organs8 attachment to the spinal cord as well as the attachment to the bladder and rectum. Once this is done, the entire organ set can be pulled out in one piece and dissected 'or 'urther investigation. v. 3uring this dissection, the various organs are e<amined and weighed and tissue samples are ta+en. %hese samples ta+e the 'orm o' PslicesP that can be viewed under a microscope. MaMor blood vessels are also bisected and e<amined. vi. %he e<aminer opens the stomach and e<amines and weighs the contents. %his can sometimes be help'ul in 'iguring out the time o' death. vii. %he e<aminer ma+es a cut with a scalpel 'rom behind one ear, across the 'orehead, to the other ear and around. %he cut is divided, and the scalp is pulled away 'rom the s+ull in two 'laps. %he 'ront 'lap goes over the patients 'ace and the rear 'lap over the bac+ o' the nec+. viii. %he s+ull is cut with an electric saw to create a PcapP that can be pried o'', e<posing the brain. ?hen the cap is pulled o'', the dura remains attached to the bottom o' the s+ull cap. %he brain is now e<posed. %he brain8s connection to the spinal cord and tentorium are severed7 brain is easily li'ted out o' the s+ull 'or e<amination. i<. %hroughout this whole process, the medical e<aminer is loo+ing 'or evidence o' trauma or other indications o' the cause o' death. %he process varies based on the nature o' the case and is incredibly detailed -- the 'orensic pathologist has to adhere to an intricate, in-depth process to ensure the proper collection and documentation o' evidence. <. Once the process is done, the organs are either put bac+ into the body or incinerated. ?hile the chest 'laps are closed and sewn bac+ together, and the s+ull cap is put bac+ in place and held there by closing and sewing the scalp. #;uipment > Dear 5sed: .. #;uipment 5sed in #<amination /. $one saw - used to cut through bone or s+ull B. $read+ni'e - used to shave slices o'' o' organs 'or e<amination H. #nterotome - special scissors used to open the intestines

C. !agedorn needle - a heavy needle used to sew up the body a'ter e<amination J. !ammer with hoo+ - used to pull s+ull cap o'' o' s+ull K. ,ib cutter - special shears used to cut through the ribs L. 6calpel - li+e a surgeon8s scalpel but with largest blade possible 'or ma+ing long deep cuts or scraping away tissue I. 6cissors - used 'or opening hollow organs and cutting vessels E. 6+ull chisel - used 'or helping to care'ully pry the s+ull cap o'' /F. 6try+er saw - the electric saw used to cut through the s+ull to remove the brain //. %oothed 'orceps - used to pic+ up heavy organs $. Dears ?orn by %echnicians 3uring .utopsy /. 6crub suits B. Downs H. Dloves (two pair) C. 6hoe covers J. *lear plastic 'ace shield Bio%#y . biopsy is the removal o' a small piece o' tissue 'or laboratory e<amination while the person is alive. O'ten times a biopsy is re;uested to test i' the patient is su''ering 'rom cancer o' the certain part o' the body, where a biopsy will be done. %ypes .ccording to #;uipment 5sed: a) 4eedle (percutaneous) $iopsy process o' removing tissues using a syringe b) Open $iopsy: procedure that uses anesthesia c) *losed $iopsy: small cut is made so that a camera-li+e instrument can be inserted which will guide the surgeon to the right place to ta+e the sample. Organ $iopsies: /. .bdominal wall 'at pad biopsy B. $iopsy o' the biliary tract H. $ladder biopsy C. $one lesion biopsy J. $one marrow biopsy K. $reast biopsy L. *arpal tunnel biopsy I. *horionic villus biopsy E. *old cone biopsy /F. *olposcopy-directed biopsy //. #ndometrial biopsy /B. Dum biopsy /H. "iver biopsy /C. "ung needle biopsy /J. "ymph node biopsy /K. Mediastinoscopy with biopsy /L. Muscle biopsy /I. Myocardial biopsy /E. 4asal mucosal biopsy BF. 4erve biopsy B/. Open lung biopsy BB. Open pleural biopsy BH. Oropharyn< lesion biopsy BC. leural needle biopsy

BJ. olyps biopsy BK. ,ectal biopsy BL. ,enal biopsy BI. 6alivary gland biopsy BE. 6+in lesion biopsy HF. 6+inny-needle biopsy H/. 6ynovial biopsy HB. %esticular biopsy HH. %ongue biopsy HC. 5pper airway biopsy HJ. 5reteral retrograde brush biopsy cytology rocedure: %he procedure o' a biopsy di''ers depending on the organ that is being biopsied. !owever a commonality among the biopsies, is that a needle is inserted into the organ and tissue is ta+en. Once the tissue is ta+en, it is sent to the lab 'or analysis, to determine the e<tent o' the disease. H and E &Hea"ato:ylin and Ea#in' Stain 2nvolves application o' hemalum, which is a comple< 'ormed 'rom aluminum ions > o<idi)ed haemato<ylin. %he nuclear staining is 'ollowed by counterstaining with an a;ueous or alcoholic solution o' eosin &, which colors other eosinophilic structures in various shades o' red, pin+ and orange. =Most commonly used stain in histopathology. !aemato<ylin 6olutions: !aemato<ylin stains are commonly employed 'or histologic studies, o'ten employed to color the nuclei o' cells (and a 'ew other obMects, such as +eratohyalin granules) blue. %he mordants used to demonstrate nuclear and cytoplasmic structures are alum and iron, 'orming la+es or colored comple<es (dye-mordant-tissue comple<es), the color o' which will depend on the salt used. .luminium salt la+es are usually colored blue-white while 'erric salt la+es are colored blue-blac+. %he three main alum haemato<ylin solutions employed are #hrlichs haemato<ylin, !arriss haemato<ylin and Mayers haemato<ylin. %he name haemalum is pre'erable to [haemato<ylin\ 'or these solutions because haematein, a product o' o<idation o' haemato<ylin, is the compound that combines with aluminium ions to 'orm the active dye-metal comple<. .lum haemato<ylin solutions impart to the nuclei o' cells a light transparent red stain which rapidly turns blue on e<posure to any neutral or al+aline li;uid. .lum or potassium aluminium sul'ate used as the mordant usually dissociates in an al+aline solution, combining with O!- o' water to 'orm insoluble aluminium hydro<ide. 2n the presence o' e<cess acid, aluminium hydro<ide cannot be 'ormed thus 'ailure o' aluminium haemato<ylin dye-la+e to 'orm, due to lac+ o' O!- ions. !ence, acid solutions o' alum haemato<ylin become red. 3uring staining alum haemato<ylin stained

sections are usually passed on to a neutral or al+aline solution (e.g. hard tap water or /S ammonium hydro<ide) in order to neutrali)e the acid and 'orm an insoluble blue aluminium haematin comple<. rocedure is +nown as blueing. 6taining o' nuclei by hemalum does not re;uire the presence o' 34. and is probably due to binding o' the dye-metal comple< to arginine-rich basic nucleoproteins such as histones. %he mechanism is di''erent 'rom that o' nuclear staining by basic (cationic) dyes such as thionine or toluidine blue. 6taining by basic dyes is prevented by chemical or en)ymatic e<traction o' nucleic acids. #<traction does not prevent staining o' nuclei by hemalum. #osin 6olutions: #osin is a 'luorescent red dye resulting 'rom the action o' bromine on 'luorescein. 2t can be used to stain cytoplasm, collagen and muscle 'ibers 'or e<amination under the microscope. 6tructures that stain readily with eosin are termed eosinophilic. #osin is most o'ten used as a counterstain to haemato<ylin in !># (haemato<ylin and eosin) staining. #osin stains red blood cells intensely red. #osin is an acidic dye and shows up in the basic parts o' the cell, ie the cytoplasm. -or staining, eosin & is typically used in concentrations o' / to J percent weight by volume, dissolved in water or ethanol. -or prevention o' mold growth in a;ueous solutions, thymol is sometimes added. . small concentration (F.J percent) o' acetic acid usually gives a deeper red stain to the tissue. Other colors, e.g. yellow and brown, can be present in the sample7 they are caused by intrinsic pigments, e.g. melanin. #;uipment 5sed in the !istopathology 6ection: Ti##ue 2roce##or is used to remove water 'rom tissues and this is replaced using a medium that solidi'ies to allow thin sections to be cut. Ti##ue cho%%er is used 'or cutting 'resh or 'i<ed tissue 'or metabolic e<periments. Microto"e is used 'or the cutting o' e<tremely thin slices o' material. Microtomy is the science 'or the preparation o' thin sections 'or materials li+e minerals, bones, and teeth. 5sing microtome, the material can be sectioned as thin as human hair. Slide #tainer is used 'or laboratory testing o' blood smears bone marrow and other bodily 'luids. Co0er #li%%er is an automated glass coverslipping instrument, used 'or highthroughput coverslipping o' several hundred slides. 0-24# 4##3"# .6 2,.%# $2O 6& (-4.$)investigates super'icial (Must under the s+in) lumps or masses.

*ytologist- cells o' tissues (*ytotechnicianM% who preps slides.) ;! Micro(iology: Microorganisms. (virology, bacteriology, mycology.) Microscopic, unicellular, and cellcluster organisms. %his includes eu+aryotes such as 'ungi and protists, and pro+aryotes. Niruses and prions, though not strictly classed as living organisms, are also studied. %ypically includes the study o' the immune system, or 2mmunology. Denerally, immune systems interact with pathogenic microbes7 these two disciplines o'ten intersect. 0Dram 6tain, culture and sensitivity, .-$ stain (acid-'ast bacilli) =gram stain: Acocci bacteria -bacilli bacteria =.-$: mycobacterium =c'u: colony 'orming units: measure o' viable bacterial or 'ungal numbers Microbiology is researched actively, and the 'ield is advancing continually. 2t is estimated only about one percent o' all o' the microbe species on #arth have been studied. .lthough microbes were directly observed over three hundred years ago, the 'ield o' microbiology can be said to be in its in'ancy relative to older biological disciplines such as )oology and botany. Bene,it# Microbes associated with various human illnesses, many microbes are also responsible 'or numerous bene'icial processes such as industrial 'ermentation (e.g. the production o' alcohol, vinegar and dairy products), antibiotic production and as vehicles 'or cloning in more comple< organisms such as plants. 6cientists have also e<ploited their +nowledge o' microbes to produce biotechnologically important en)ymes such as %a; polymerase, reporter genes 'or use in other genetic systems and novel molecular biology techni;ues such as the yeast two-hybrid system. $acteria can be used 'or the industrial production o' amino acids. orynebacterium glutamicum is one o' the most important bacterial species with an annual production o' more than two million tons o' amino acids, mainly "-glutamate and "-lysine. . variety o' biopolymers, such as polysaccharides, polyesters, and polyamides, are produced by microorganisms. Microorganisms are used 'or the biotechnological production o' biopolymers with tailored properties suitable 'or high-value medical application such as tissue engineering and drug delivery. Microorganisms are used 'or the biosynthesis o' <anthan, alginate, cellulose, cyanophycin, poly(gammaglutamic acid), levan, hyaluronic acid, organic acids, oligosaccharides andpolysaccharide, and polyhydro<yal+anoates.

Microorganisms are bene'icial 'or microbial biodegradation or bioremediation o' domestic, agricultural and industrial wastes and subsur'ace pollution in soils, sediments and marine environments. %he ability o' each microorganism to degrade to<ic waste depends on the nature o' each contaminant. 6ince sites typically have multiple pollutant types, the most e''ective approach to microbial biodegradation is to use a mi<ture o' bacterial species and strains, each speci'ic to thebiodegradation o' one or more types o' contaminants. %here are also various claims concerning the contributions to human and animal health by consuming probiotics (bacteria potentially bene'icial to the digestive system) and(or prebiotics (substances consumed to promote the growth o' probiotic microorganisms). ,ecent research has suggested that microorganisms could be use'ul in the treatment o' cancer. Narious strains o' nonpathogenic clostridia can in'iltrate and replicate within solid tumors. *lostridial vectors can be sa'ely administered and their potential to deliver therapeutic proteins has been demonstrated in a variety o' preclinical models. #;uipment: /. .gar plate: a etri dish that contains a growth medium (typically agar plus nutrients) used to culture microorganisms or small plants li+e the moss !hyscomitrella patens. B. .ir displacement pipettes: istondriven7 tools to handle volumes o' li;uid in the microliter scale and are the most accurate and precise pipettes. %hey are more commonly used in biology and biochemistry, but are also used by chemists, though less commonly due to the ris+ o' solvent damage to the pipettes and tips7 micropipettes. H. .ntibiotic discs: antibioticimpregnated paper discs used in 1irby-$auer antibiotic testing. C. .utoclave: instrument used to sterili)e e;uipment and supplies by subMecting them to high pressure saturated steam at /B/ V* 'or around /JGBF minutes depending on the si)e o' the load and the contents. J. .utoclave tape: adhesive tape used to heat with steam (high pressure) to sterili)e to indicate whether a speci'ic temperature has been reached. .utoclave tape wor+s by changing color a'ter e<posure to temperatures commonly used in sterili)ation processes, typically /B/V* in a steam autoclave. K. $er+e'eld: bacterial water 'ilter used in homes, microbiological laboratories, and in the 'ield.

L.

*ollodion bag: a membrane used to 'ilter or concentrate substances, o'ten proteins, using pressure. %a+es the 'orm o' a small 'inger shaped receptacle hoo+ed up to a positive pressure pump. I. *olony counter: instrument used to count bacteria colonies or other microorganisms growing on an agar plate. E. 3urham tubes: used in microbiology to detect production o' gas by microorganisms. /F. 2ncubator: device used to grow and maintain microbiological cultures or cell cultures. //. 2noculation loop" smear loop, inoculation wand or microstreaker7 is a simple tool used mainly by microbiologists to retrieve an inoculum 'rom a culture o' microorganisms. /B. "aboratory water bath: tool used to maintain a very stable temperature li+e an incubator. /H. Microscope: instrument used to see obMects that are too small 'or the na+ed eye. /C. etri dish: etri plate or cell culture dish7 is a shallow glass or plastic cylindrical lidded dish that biologists use to culture cells or small moss plants. /J. ipette: laboratory tool used to transport a measured volume o' li;uid. MYCOLO7Y: study or branch o' botany 7 'ocuses on 'ungi. *an be bro+en down into a number o' other areas: medical or clinical mycology, mold mycology, etc. urpose o' the study: understand more about the characteristics and growth patterns o' 'ungi, and how they may a''ect humans 'or the good or the bad. 6ome may help people, others can be e<tremely harm'ul. 2n essence, mycology is the root o' many di''erent types o' studies, including the study o' beer, wine, cheeses, medicine, and a number o' other things. 5nderstanding how best to use 'ungi when ma+ing these products is di''icult without a 'irm grasp into what 'ungi are. 2denti'ied as many as appro<imately BFF,FFF species o' 'ungi, though not all are subMect to e<tensive study, and Q/S is +nown to cause problems 'or humans. %hose that do are sometimes simple nuisances, such as the 'ungus responsible 'or athlete8s 'oot (#inea pedis). Others may cause more serious problems, such as tumors, that re;uire immediate medical attention. #ven seemingly harmless 'ungi could, i' le't untreated, cause problems that lead to other types o' in'ection. %here'ore, medical mycology studies these 'ungi and attempts to discover better treatment methods. Medical mycology also may see+ to determine what types o' 'ungi may be bene'icial 'or human use, either as a 'ood source or as a

source o' medicine. -or e<ample, even early humans understood the importance o' studying some types o' 'ungi, and learned early on that some have desirable traits, such as yeast, which is used in breads. &east is also used as an antibiotic drug 'or those su''ering various in'ections. 2n some cases, where 'ungus is thought to be the source o' a health problem in humans or animals, mycologists could help determine the source o' the contamination. 2n such cases, the mycologist loo+s 'or a 'ungus and then tries to determine the species. 3epending on what is +nown about the 'ungus, this may also help isolate the source, which is important 'or eliminating the problem. %his has application not only inside o' people8s homes, but also in protecting entire 'ood systems. ) .*23--.6% $.*2""2 6%.24 (.-$ 6%.24) One o' several types o' tests used to identi'y di''erent bacteria. .-$ is used primarily to identi'y bacteria in the genus Mycobacterium, such as Mycobacterium tuberculosis. rocess includes staining the specimen and then trying to wash out that stain by applying an acid. .-$ smear is used to loo+ 'or .-$ in a sample 'rom the site o' suspected in'ection. -or the smear, the sample is spread thinly onto a glass slide, treated with a special stain, and e<amined under a microscope. %his is a relatively ;uic+ way to determine i' an in'ection may be due to one o' the acid-'ast bacilli, the most common o' which is M$ tuberculosis. .n .-$ smear may be done in conMunction with a molecular test 'or %$ termed nucleic acid ampli'ication test (4..%). %hese tests are not diagnostic, but they can provide a presumptive diagnosis, which can aid in the decision o' whether to begin treatment be'ore culture results are available. .-$ cultures are used to diagnose active M$ tuberculosis in'ections, in'ections due to another member o' the Mycobacterium 'amily, or to determine whether %$-li+e symptoms are due to another cause. %hey are used to help determine whether the %$ is con'ined to the lungs (pulmonary disease) or has spread to organs outside the lungs (e<trapulmonary disease). .-$ cultures can also be used to monitor the e''ectiveness o' treatment and can help determine when a patient is no longer in'ectious. ,esults: -.-$ smear @ no in'ection is present, that symptoms are caused by something other than mycobacteria, or that the mycobacteria were not present in su''icient numbers to be seen under the microscope. 5sually at least three samples are collected to increase the probability that the organisms will be detected. 4evertheless, i' .-$ smears are negative and there is still a strong suspicion o' a mycobacterial in'ection, then additional samples may be collected and tested on di''erent days. . smear negative sample may

still grow mycobacteria since the culture media allows low numbers o' bacteria to multiply and be detected. A.-$ smears indicate a probable mycobacterial in'ection. !owever, a culture must be per'ormed to con'irm a diagnosis. .-$ testing is ordered when: 6omeone has symptoms that suggest pulmonary %$ or other mycobacterial lung in'ection, such as: o "ingering, chronic cough that produces phlegm or sputum, sometimes with bloody strea+s o -ever, chills o 4ight sweats o 5ne<plained weight loss o ?ea+ness o *hest pain 6omeone has symptoms associated with %$ or other mycobacterial in'ection outside o' the lungs (e<trapulmonary). 6ymptoms vary depending on area o' the body a''ected. #<amples include bac+ pain and paralysis (spinal %$), wea+ness due to anemia (%$ inbone marrow), altered mental state, headache, > coma (%$ meningitis), Moint pain or abdominal pain. %$ screen test is positive and the person is at increased ris+ 'or active disease and(or has characteristic lung involvement as shown by U-ray. 6omeone has been in close contact with a person who has been diagnosed with %$, and the e<posed person either has symptoms or has a condition or disease that puts them at a much higher ris+ o' contracting the disease, such as!2N(.236. (%hose with .236 are more li+ely than other a''ected patients to have e<trapulmonary %$ with a 'ew, vague symptoms.) 6omeone is being treated 'or %$7 .-$ smears and cultures are usually ordered at intervals, both when the doctor is evaluating the e''ectiveness o' treatment and when he is attempting to determine whether or not a person is still in'ectious. ) D,.M 6%.2424D Method o' di''erentiating bacterial species into two large groups (Dram-positive and Dramnegative). %he Dram stain is almost always the 'irst step in the identi'ication o' a bacterial organism, and is the de'ault stain per'ormed by laboratories over a sample when no speci'ic culture is re'erred. Method named a'ter inventor, 3anish scientist !ans *hristian Dram. Dram devised his techni;ue not 'or the purpose o' distinguishing one group o' bacteria 'rom another but to enable bacteria to be seen more readily in stained sections o' lung tissue. ublished method in /IIC. Dram stains are per'ormed on body 'luid or biopsy when in'ection is suspected.

Dram stains yield results much more ;uic+ly than culture, and is especially important when in'ection would ma+e an important di''erence in the patient8s treatment and prognosis. . Dram stain and culture o' the material 'rom an in'ected site are the most commonly per'ormed microbiology tests used to identi'y the cause o' an in'ection. O'ten, detecting the presence o' microorganisms and determining whether an in'ection is caused by an organism that is Dram-positive or Dram-negative will be su''icient to allow a doctor to prescribe treatment with an appropriate antibiotic while waiting 'or more speci'ic tests, such as a culture, to be completed. .bsence or presence o' white blood cells in the Dram stain can help establish that an ade;uate sample was obtained as white blood cells are 're;uently present with an in'ection. . Dram stain may also be per'ormed as part o' the evaluation o' a culture. ?hen bacteria grow on(in a nutrient media in the laboratory, a Dram stain is per'ormed to help determine the type o' bacteria present and to help determine what other tests may need to be per'ormed to de'initively identi'y the cause o' in'ection. rocedure: .. 6lant *ultures - . culture made on the slanting sur'ace o' a solidi'ied medium in a test tube that has been tilted to provide a greater area 'or growth. .lso called slope culture /. repare and heat-'i< smears. B. repare the smears o' 6. epidermidis and 4. sicca on a second slide. !eat-'i<. H. 6tain the slides as 'ollows: a. -lood the crystal violet 'or one minute. b. our o'' e<cess dye and wash gently in tap water and drain the slide against a paper towel. c. #<pose the smears to Dram8s iodine 'or one minute by washing with iodine, then adding more iodine and leaving it on the smear until the minute is over. d. ?ash with tap water and drain care'ully. (3o not blot.) e. ?ash with EJS alcohol 'or HF seconds. '. ?ash with tap water at the end o' the HF seconds to stop the decolori)ation. 3rain. g. *ounterstain with F.BJS sa'ranin 'or HF seconds. h. ?ash, drain, blot, and e<amine under oil. i. 3raw the cells showing morphology, grouping, and relative si)es. *olor a 'ew o' the cells o' each bacterial species to show the Dram reaction. M. 6ave these slides and the ones 'rom parts $ > * o' this e<ercise to use at the ne<t lab period. $. $roth *ultures - bacteria or any other

growing microorganism in li;uid nutritional medium (broth) in test tube /. 6mear made 'rom the broth will be a thin smear and nearly invisible to the na+ed eye even a'ter staining, may be advisable to draw a ring with a 'elt pen on the underside o' the slide to mar+ the area in which the broth smear will be made. ?hen ma+ing a smear 'rom broth: do not add a drop o' water to the slide. B. !ear-'i< the smears, Dram stain them with the above procedure, and e<amine them. ?hen 'ocusing the broth smear use the techni;ue suggested 'or thin smears. H. *ompare the appearance o' the cells in the two smears. . negative Dram stain is o'ten reported as Pno organism seen.P %his may mean that there is no bacterial in'ection present or that there were not enough microorganisms present in the sample to be seen with the stain under a microscope. ositive Dram stain results usually include a description o' what was seen on the slide. %his typically includes whether the bacteria are Dram-positive (purple) or Dram-negative (pin+) as well as their shape round (cocci) or rods (bacilli). 6ometimes si)e, relative ;uantity, and grouping o' the bacteria, i' relevant, are also reported. 2' there are bacteria present within other cells (intracellular), this will also be noted as well as7 'or e<ample, the presence o' red blood cells or white blood cells. %his in'ormation, along with signs and symptoms and other clinical 'indings, will help the doctor decide which treatment may be most e''ective, o'ten be'ore culture results are available. !owever, Dram stain results are usually considered preliminary, and results o' a culture and(or other tests such as antigen and antibody testing 'or particular types o' bacteria are necessary to con'irm a diagnosis. 6ometimes, susceptibility testing is necessary to determine which antimicrobial will be most e''ective in treating the in'ection. 6ome body sites are generally sterile, such as the blood and cerebrospinal 'luid. $acteria will not usually be present in these sites when there is not a bacterial in'ection. %hey may initially be present in low numbers with an in'ection, and a sample may re;uire e<tra processing in order to concentrate the bacteria so that they can be detected by a Dram stain. Other body 'luids and sites, such as sputum or s+in, typically have cells and normal 'lora present in addition to any bacteria that are causing an in'ection. Dram stains on these types o' samples re;uire care'ul e<amination by a trained laboratorian to determine which cells may be normal and which may be due to an in'ection. ) *5"%5,#: Microbiological culture, or microbial culture7 method o'

multiplying microbial organisms by letting them reproduce in predetermined culture media under controlled laboratory conditions. Microbial cultures are used to determine the type o' organism, its abundance in the sample being tested, or both. 2t is one o' the primary diagnostic methods o' microbiology and used as a tool to determine the cause o' in'ectious disease by letting the agent multiply in a predetermined medium. Microbial cultures are 'oundational and basic diagnostic methods used e<tensively as a research tool in molecular biology. 2t is o'ten essential to isolate a pure culture o' microorganisms. (. pure (a%enic) culture is a population o' cells or multicellular organisms growing in the absence o' other species or types. . pure culture may originate 'rom a single cell or single organism, in which case the cells are genetic clones o' one another.) Culture o, Micro3organi#"# $acteria, roto)oa and .lgae: *an be success'ully cultured both on solid (agar) media and in li;uid (broth). .ctinomycetes: Optically clear plastic containers 'acilitate the observation o' the 'ascinating behaviour o' these cells especially under the conversion 'rom amoeboid, single cell 'orm to that o' a multicellular organism with the necessary di''erentiation o' di''erent 'unctional cell types. -ungi: 3epending on the application these versatile organisms can be cultured to harvest their secreted products e.g. antibiotics or to study other aspects o' the organisms in their own right, either in li;uid culture o' on an agar sur'ace. %he procedures to use in culturing micooganisms depend on which types o' organisms you wish to study. Culturing Bacteria: Drowth Media: /(/F-strength %rypticase 6oy .gar (%6.) Media: 2ngredients: B g %rypticase 6oy .gar L.J g $acto .gar JFF ml distilled water Mi< the ingredients, autoclave 'or BF minutes, and pour into sterile petri dishes. late out the bacteria using a /F-L dilution starting with J g dry weight o' the compost in CJ ml o' an autoclaved .FKM 4a! OC(4a!B OC bu''er o' L.K p!. (appro<imately C:/ dibasic:monobasic). ut this 'irst dilution in a blender 'or CF sec. at high speed. er'orm serial dilutions to /F-L and add F!/ ml o' the dilution per plate. 2ncubate at BI* 'or C days. *ount the colonies as colonies per unit a'ter C days. repare slides o' speci'ic colonies the same day.

Culturing Actino"ycete# Drowth Media: /(JF-strength %6. oly $ 2ngredients: F.C g %ypticase 6oy .gar /F.F g $acto .gar JFF ml distilled water /F mg olymi<in $ in /F ml LFS #thanol Mi< the 'irst H ingredients, autoclave 'or BF minutes, and cool to room temperature. .dd the antibiotic and pour into sterile petri dishes. late out the actinomycetes using a /FL dilution starting with J g dry weight o' compost in CJ ml o' the autoclaved bu''er. ut this 'irst dilution in a blender at high speed 'or CF sec. er'orm serial dilutions to /F-L and add F.l ml o' the 'inal dilution to each plate. 2ncubate the plates at BI* 'or /C days. %a+e counts and samples o' actinomycetes colonies a'ter /C days. Many o' the colonies will loo+ powdery white. !owever, some may ta+e on a rough appearance and produce a variety o' pigments. 4ote: 2' you are comparing mesophilic compost to thermophilic compost, you should prepare double the usual number o' plates so that you can incubate plates at both BI* and JF*. Culturing Fungi Drowth Media: /(H-strength 3., 2ngredients: K.J g otato 3e<trose .gar J.F g $acto .gar JFF ml distilled water /J mg ,i'ampicin in /F ml Methanol /J mg enicillin D in /F ml LFS #thanol Mi< the 'irst H ingredients, autoclave 'or BF min. and cool to room temperature. .dd the antibiotics and pour into sterile petri dishes. late out the 'ungi using a /F-C dilution starting with Jg dry weight o' compost in CJ ml o' the autoclaved phosphate bu''er. ut this 'irst dilution in a blender at high speed 'or CF sec. er'orm serials dilutions to /F-C and add F.l ml o' the 'inal dilution to each plate. 2ncubate the plates at BI* 'or H days. %a+e counts and samples o' 'ungal colonies at H days. (.gar, or agarose gel, used as a medium 'or gelling the microbial culture. 2t is a gelatinous substance derived 'rom seaweed. . cheap substitute 'or agar is guar gum, which can be used 'or the isolation and maintenance o' thermophiles.) 06#462%2N2%&: 6#462%2N# @ a clear, circular PhaloP (technically +nown as a Ppla;ue,P or )one o' inhibition) will appear around the antibiotic dis+, indicating an absence o' bacteria. .ntibiotic has inhibited their growth and(or +illed them@the particular antibiotic should be e''ective against the in'ection the person has7

chec+s to see what +ind o' medicine, such as an antibiotic, will wor+ best to treat the illness or in'ection 5IROLO7Y Nirology is a branch o' the sciences which 'ocuses on the study o' viruses and organisms which behave li+e viruses, such as prions and viroids. One o' the primary goals o' virology is classi'ication, in which viruses are studied to determine what they are and how they wor+. *lassi'ication can be used to determine that various viruses are related to each other, and that they may there'ore wor+ in the same way, or be vulnerable to the same antiviral drugs. $eing able to classi'y viruses also allows researchers to determine i' the virus has been seen be'ore, > lin+ the viruses they 'ind with e<isting studies and in'ormation. Nirologists also are concerned with the structure o' viruses, and the way in which viruses wor+. %hough not considered living organisms, viruses can be ;uite comple<, and they have adapted a number o' clever tric+s, li+e hiMac+ing cells and getting them to reproduce the virus or tric+ing the body into thin+ing that a viral agent is not an unwanted invader. 5nderstanding how these organisms wor+ @ important part o' developing methods to eradicate them. Niral diseases, the outcome o' viral in'ection, are also o' interest to virologists, along with modes o' transmission and related topics. ?hen outbrea+s o' viruses occur, researchers conduct research to determine where the virus came 'rom, how it can be treated, what the symptoms are, and how additional in'ections might be prevented. Nirologists also trac+ long term trends, such as changes in viral 34., or alterations in immunity levels in populations which are at ris+ o' in'ection. Microbiologist <! Clinical Micro#co%y: (body 'luids not blood) 5rine and other bodily 'luids. %his section deals with the study o' the gross, chemical and microscopic analysis o' di''erent body 'luids other than blood. (*erebrospinal 'luid (*6-)- spinal cavity7 6ynovial 'luid- +nees7 leural 'luid- lungs) ) #--562O46: ?hen 'luids accumulate inside a cavity they are re'erred to as e''usions. #''usions occur in the pleural, pericardial, and peritoneal cavities o' the body. 6ince it is not natural 'or 'luids to accumulate in these spaces it is important that they be tested by a laboratory in order to determine the cause o' the accumulation. #''usions are classi'ied as either transudates or e<udates. TRANS$-ATES ANE=$-ATES: *lassi'ying a 'luid as a transudate or an e<udate can help determine the disease process causing 'luid to accumulate and

proceed with subse;uent treatment. .ll organs in the body have their own linings that help to protect the organ and are 'iltered by the lymphatic system. 2n order 'or these linings to wor+ properly they must be permeable and allow 'or the trans'er o' 'luids, proteins, and other metabolites between them. Tran#udate# .ccumulates in cavities due to a mal'unction o' the 'iltering membranes o' cavity linings E:udate# .ccumulates in cavities due to the presence o' 'oreign materials such as bacteria, viruses, parasites, 'ungi, and tumor cells -orms as a result o' leu+ocytes, 'oreign material and their metabolites 'illing the cavity.

5sually 'ound in conditions such as liver disease, pancreatic disease, and congestive heart 'ailure. *an be thought as a result o' mechanical process.

*an be thought as a result o' in'lammatory process. 5rine: "i;uid waste product o' the body secreted by the +idneys and e<creted through the urethra. %wo uni;ue characteristics o' urine: ,eadily available and easily collected !as in'o on bodys maMor metabolic 'unctions *omposition: . metabolic waste product produced in *onsists o' urea and other organic and inorganic e.g. creatinine and uric compounds dissolved in water e.g. chlorine, sodium and
acid potassium
the liver 'rom the brea+down o' protein and amino acids

) 5,24."&626 (5.): group o' chemical and microscopic tests(e<aminations done on a urine sample. 5rine contains a range o' substances that vary with what is introduced into the body. .side 'rom water, urine contains an assortment o' inorganic salts and organic compounds, including proteins, hormones, and a wide range o' metabolites. %he macroscopic, chemical and microscopic e<aminations o' urine provide initial valuable diagnostic in'ormation concerning metabolic dys'unctions o' both renal and non-renal origin. 3etect byproducts o' normal and abnormal metabolism, cells, cellular 'ragments, and bacteria in urine. Many disorders can be diagnosed in their early stages by detecting abnormalities in the urine. .bnormalities include increased concentrations o' constituents that are not usually 'ound in signi'icant ;uantities in the urine, such as:

glucose, protein, red blood cells, white blood cells, crystals, and bacteria. #]52 M#4%(M.%#,2."6: Microscope *linical *entri'uge ,e'ractometer Multiple reagent dipstic+s 6 #*2M#4 *O""#*%2O4 .43 !.43"24D: /. 6pecimen must be collected in a clean dry, disposable container B. %he container must be labeled with the patients name together with the date and time o' collection H. 6pecimen must be delivered to the laboratory and tested within one hour =specimen that cannot be delivered within one hour should be re'rigerated or preserved chemically 5,24# 6 #*2M#46: ,.43OM 6 #*2M#4: most commonly received due to ease o' collection A lac+ o' inconvenience. 5se'ul in routine screening tests to detect obvious abnormalities. !owever, it may also produce erroneous results caused by dietary inta+e or physical activity Must prior to the collection o' the specimen. -2,6% MO,424D 6 #*2M#4: ideal screening specimen. *oncentrated, assuring detection o' chemicals and 'ormed elements that may not be present in a dilute random specimen. B-!O5, O6% ,.432." 6 #*2M#4: patient instructed to void shortly be'ore consuming a routine meal and to collect a specimen B hours a'ter eating. 6pecimen is then tested 'or glucose and results are used primarily 'or monitoring insulin therapy in persons with diabetes. -.6%24D(6#*O43 MO,424D 6 #*2M#4 G the second voided specimen a'ter a period o' 'asting (urine voided a'ter 'irst morning specimen). %his specimen will not contain any metabolites 'rom 'ood ingested prior to the beginning o' the 'asting period and is recommended 'or glucose monitoring. M23 6%,#.M [*"#.4 *.%*!\ 6 #*2M#4 G pre'erred type o' specimen 'or culture and sensitivity testing because o' the reduced incidence o' cellular and microbial contamination. *.%!#%#, *O""#*%2O4 6 #*2M#4 G urine collected 'rom a catheter, a hollow tube passing to the urethra into the bladder. %2M#3 6 #*2M#4 G collection o' all urine in a span o' BC hours or as prescribed. 65 ,.- 5$2* .6 2,.%2O4 6 #*2M#4 G collected 'rom a non-ambulatory patient who cannot be catheteri)ed or where there are concerns about obtaining a sterile specimen by conventional means. 2t is collected by e<ternal introduction o' a needle into the bladder.

#32.%,2* 6 #*2M#4 G use o' a special urine collection bag that is adhered to the s+in surrounding the urethral area 'or in'ants and small children. 5,24."&626 #U.M24.%2O46: a. D,O66(M.*,O6*O 2* #U.M24.%2O46 G hysical routine e<amination o' the specimen Nolume G dependent on the amount o' water e<creted by the +idneys7 amount e<creted is determined by the bodys state o' hydration o -actors that in'luence urine volume -luid inta+e -luid loss Nariation in secretion o' anti diuretic hormone o /BFF G /JFF ml G daily urine output o More than HFFFml(day G olyuria G increase in daily urine volume due to diabetes o "ess than JFFml(day G Oliguria Gdecrease in the normal daily urine volume due to dehydration o 4octuria G increase in nocturnal e<cretion o' urine7 associated with polyuria o .nuria: stop o' urine 'low7 =obstruction and heart 'ailure Odor G 6mell o' urine a''ected by 'ood. 6ome 'oods and beverages that contribute to odor: alcohol, co''ee, asparagus, sa''ron, tur+ey, and onion. 6picy 'oods can have a similar e''ect, as compounds are not bro+en down by the +idneys and pass through the body in the urine. o -aint(.romatic odor: normal o .mmonia-li+e odor: caused by brea+down o' urea7 indicates presence o' a urea-splitting bacteria o -oul(o''ensive odor: old specimen or presence o' pus or in'lammation o 6weet odor: presence o' glucose o Maple syrup-li+e: Maple 6yrup urine disease (M653), a serious metabolic de'ect o -ruity odor: presence o' +etones o ungent odor: ingestion o' some 'oods whose compounds are not bro+en down by the +idneys *O"O, o *olorless G very dilute urine o "ight to medium yellow G normal (yellowing to light orange is caused by removal o' e<cess $ Nitamins 'rom the bloodstream and certain medications) o Nery dar+ yellow G e<tremely concentrated7 caused by ,ibo'lavin7 may be indicative o' bilirubinuria (presence o' bilirubin) and(or dehydration o &ellow green G caused by bilirubin o<idi)ed to bilivedrin7 may also be

caused by consumption o' asparagus and dietary $ vitamin supplement. o Dreen G indicative o' seudomonas in'ection o ,ed to brownish red G bloody urine or hematuria (presence o' red blood cells in urine) which is potentially a sign o' bladder in'ection or carcinoma7 indicative o' hemoglobinuria (high concentrations o' hemoglobin) and myoglobinuria (presence o' myoglobin) o ,eddish brown to brown G indicative o' hemoglobinuria, myoglobinuria and presence o' methemoglobin (o<idi)ed ,$*7 abnormal hemoglobin)7 can be a symptom o' Maundice, rhabdomyolysis and Dilberts syndrome o $lac+ or dar+-colored G melanuria (caused by a melanoma) %ransparency(%urbidity o *lear, slightly cloudy G normal (in 'resh urine) =clear urine is not always normal o *loudy, opa;ue, 'locculent G abnormal robable causes: presence o' ,$*, ?$*, epithelial cells, mucus strands, numerous crystals, bacteria, lipids, semen(vaginal secretions, lymph 'luid, yeast and 'ecal contamination b. *!#M2*." .4."&26 (32 6%2*1 .4."&26) G 3ipstic+s or reagent stic+s consist o' various pads containing chemical ingredients which provide a color change when a particular analyte is present in urine. %his color change is converted to a semi-;uantitative result 'or the analyte in ;uestion. 2t is used to give results in p!, protein, glucose, +etones, bilirubin and proteins containing a heme group. c. M2*,O6*O 2* .4."&626 G 3one to identi'y insoluble constituents o' urine and correlate the physical and chemical urinalysis results with the microscopic observations and recogni)e discrepancies. 6 #*2-2* D,.N2%& G measure o' the solute concentration and density o' urine compared to pure water7 detects possible dehydration o 4ormal values ('or adults) are between /.F/F to /.FBI 2ncreased 6 D,.N G hypersthenuria (high concentration o' solutes in urine) 3ecreased 6 D,.N G hyposthenuria (low concentration o' solutes in urine) p! "#N#" o .cidic G p! level below L o 4eutral G p! level is e;ual to L o $asic G p! level is above L D"5*O6# "#N#" o 4ormal value urine glucose G /HFmg(BC glycogen hours
Only carbohydrate to be directly used 'or energy or stored as

=glucose levels 'luctuate7 even a normal person may have glucosuria (presence o' glucose in urine) when a meal with high glucose content is eaten =urine that is to be tested 'or glucose levels must be collected a'ter 'asting o .bnormal glucose levels may be indicative o' the 'ollowing: 3iabetes Mellitus !ypoglycemia 2mpaired tubular re absorption *entral 4ervous 6ystem damage %hyroid disorders ,O%#24 G normal urine contains very little protein, usually less than /F mg(dl or /FFmg(BChours is e<creted. o 2ncrease in protein levels may be diagnosed as: Multiple myeloma re-eclempsia 3iabetic nephropathy $2"2,5$24 G provides indication(detection o' liver disease, hepatitis, cirrhosis, gallbladder disease and cancer. 1#%O4#6 =4ormally, measurable amounts o' +etones do not appear in urine because all o' the metaboli)ed 'at is completely bro+en down into *OB and !BO =*linical reasons 'or this increased 'at metabolism include the inability to metaboli)e carbohydrates (as occurs in diabetes), increased loss o' carbohydrates 'orm vomiting, and inade;uate inta+e o' carbohydrates associated with starvation and weight reduction resence o' ,$* and ?$* in ! - (!igh power 'ield) o "ess than J ?$*(! - G normal More than J ?$*(! - G yuria, indicating the presence o' an in'lammatory process somewhere in the urinary tract7 yuria is caused by 5.%.2. and(or bacteria o 5p to J ,$*(! - G normal More than J ,$*(! - G hematuria, due to hemorrhage, in'lammation, necrosis, trauma, or neoplasia somewhere along the urinary tract o resence o' bacteria and epithelial cells: 4one, 'ew, moderate or many resence o' crystals: 4one, 'ew, moderate or many. ) 6 #,M(6#M24." -"523: Organic 'luid, may contain spermato)oa, secreted by the male se<ual glands (gonads) that can 'ertili)e 'emale ova G Most 're;uently received in *linical "aboratories 'or two primary reasons: /.) #valuation o' 'ertility cases and B.) ost vasectomy cases *OM O62%2O4:
2ntermediate products o' 'at metabolism

6permato)oa G produced in the testis and mature in the epididymis7 contributes to a small amount o' total semen volume rostate G a mil+y 'luid responsible 'or coagulation and li;ue'action o' semen *O""#*%2O4: /. 6pecimen should be collected in sterile containers 'ollowing a H-day period o' se<ual abstinence B. 5se o' condoms 'or specimen collection is not recommended 'or 'ertility testing because the condoms may contain spermicidal agents H. 6pecimen should be +ept at room temperature and delivered to the laboratory within one hour C. . 'resh semen specimen is clotted and should li;ue'y within HF minutes a'ter collection7 thus the time o' collection is essential 'or evaluation o' semen li;ue'action J. 6pecimens awaiting motility analysis : +ept at HLo* 6#M#4 .4."&626: Nolume: 4ormal is B-J ml per eMaculation Niscosity: . specimen o' normal viscosity will pour in droplets and will not appear clumped or stringy p! level: 4ormal range is L.H to I.H7 slightly al+aline 6perm count: BF million G /KF million sperm per m" Motility (microscopically e<amine undiluted sperm and determine the percentage o' sperm showing active motility): OJF-KFS within H hours ]uality: O B.F or 'air Morphology (evaluated with respect to both head and tail appearance): Q HFS abnormal 'orms

) 6&4ON2." -"523: [Moint 'luid\7 viscous 'luid 'ound in Moint cavities7 supplies nutrients to the cartilage and acts as a lubricant to sur'aces o' the 'ree moving Moints

=normal amount o' 'luid contained in the +nee cavity is less than H.Jml *O""#*%2O4: /. *ollected by needle aspiration (arthrocentesis) B. 4ormal synovial 'luid wont clot7 'luid 'rom diseased Moint may contain 'ibrinogen and clot H. H tubes routinely collected 6terile heparini)ed tube 'or microbiology .nti coagulated 'or hematology 4on anti coagulated tube 'or other tests 6&4ON2." -"523 .4."&626: *olor: 4ormal synovial 'luid appears clear and pale yellow7 =deeper yellow @ in'lammation7 greenish tinge @ bacterial in'ection %urbidity: cell count7 / number o' cells present7 mil+y 'luid may also indicate crystals Niscosity: -rom the polymeri)ation o' the hyaluronic acid7 essential 'or the proper lubrication o' Moints *ell counts: ?$* counts below BFF cells(microliter are normal7 may reach /FF,FFF cells(microliter in severe in'lammations *rystal identi'ication: rimary crystals seen in synovial 'luid are: Monosodium urate (uric acid), cases o' gout *alcium pyrophosphate, cases o' pseudogout cholesterol crystals crystals o' apatite cortico steroid crystals, 'rom drug in'ection )*#,#$,O6 24." -"523 (*6-): clear, colorless bodily 'luid in ventricles o' brain7 occupies the subarachnoid space and the ventricular system around and inside the brain and spinal cord. %he brain P'loatsP in it. 2t can be withdrawn by per'orming a spinal or lumbar puncture (the insertion o' a needle through the bac+ and into the spinal canal, permitting withdrawal o' spinal 'luid). *6- .4."&626: set o' laboratory tests that e<amine a sample o' the 'luid surrounding the brain and spinal cord. %his test may be ordered when a doctor suspects that a patient

has a condition or disease involving their *46 (*entral 4ervous 6ystem). ressure: can be measured when opening (starting) > closing ('inishing) collection. =4ormal *6- ressure is at I-/Jmm!g with the patient lying on the side and /K-HB mm!g when patient is sitting up. =2ncreased(#levated *6- ressure G cerebral blood 'low may be constricted due to tumors, in'ection, abnormal accumulation o' *6- within the brain (hydrocephalus), and bleeding. =3ecreased *6ressure G due to dehydration, shoc+, or lea+age o' *6through an opening. *olor: *lear(*olorless @ normal7 &ellow, Orange or in+ G <anthochromic (caused by ,$*s entering the *6- during bleeding)7 Dreen: bilirubin or in'ection %urbidity: *loudy(%urbid G indicate the presence o' white or red blood cells, microorganisms, or an increase in protein levels. Niscosity: 4ormal *6- @ same consistency as water. *6- that is [thic+er\ may be seen in patients with certain types o' cancers or meningitis. Dlucose: 4ormal is JF to IF mg(d" (or greater than two-thirds o' blood sugar level)=Dlucose CF mg(d" is signi'icant and occurs in bacterial and 'ungal meningitis and in malignancy. rotein: 4ormal is /J -KF mg(d"7 =!igh levels are seen in many conditions including: bacterial and 'ungal meningitis, multiple sclerosis, tumors, subarachnoid hemorrhage ,$* and ?$* count: 4ormally no red blood cells are present in the *6-. resence o' ,$* may indicate bleeding into the *6- or may indicate a [traumatic tap\ - blood that lea+ed into the *6- sample during collection. 4ormally less than J cells are present in the adult. . signi'icant increase in white blood cells in the *6- is seen with in'ection or in'lammation o' the *entral 4ervous 6ystem. Dram 6tain: direct observation o' microorganisms under microscope. 6hould have no microorganisms in *6- 'luid. 2' bacteria or 'ungi are present on a *6gram stain, then the patient has bacterial or 'ungal meningitis or encephalitis. *linical Microscopist- pericardial 'luid, *6-, saliva, sperm analysis, et cetera. >! 2ara#itology: 6tool, blood, tissue, anywhere with parasites. arasites are organisms that spend portion o' its li'e in or on the living tissue and which causes harm to the host without immediately +illing it. 4ematode 2n'ections: 4early HFF species o' parasitic worms and LF species o' proto)oa

can inhabit a human body. Many are rare and accidental7 9 EF common species. %apeworms: (*estodiasis) %aeniid: %he two species are the #aenia saginata, or bee' tapeworm and the #aenia Solium, or also +nown as the por+ tapeworm. 3iphyllobothrinid: 'ish tapeworm also lives in the intestines !oo+worm: (.ncyclostoma) &ncylostoma duodenale (Old ?orld, .sia) 'ecator americanus (4ew ?orld, .'rica) 6mall intestine, cause massive blood loss. #ggs pass out with the 'eces to contaminate the soil, where larvae emerge and molt to become in'ectious larvae that bore through the s+in o' a new host. ,oundworm: &scaris lumbricoides: lives in the lumen o' small intestine7 6oil transmitted inworm( 6eatworm: (#nterobius) (nteroblus )ermicularis: "ives in the large intestine: cecum, ileum, colon, appendi<7 %ransmitted through direct anus to mouth by 'inger contamination, 'inger to mouth contamination, airborne, retroin'ection ?hipworm: (%richuris) #richuris trichiura" $uries anterior in mucosa o' cecum7 transmitted via ingestion o' embryonated egg. (ntamoeba histolytica" one o' nine species o' intestinal amoebae7 only pathogen7 live and multiply in the gut and 'orm cysts7 passed in the 'eces and in'ect new individuals when they are consumed in contaminated water or 'ood. Most in'ections are asymptomatic, but some strains can invade the gut wall, causing severe ulceration and amoebic dysentery @ bloody stools. 2' the parasites gain access to damaged blood vessels, they may be carried to e<traintestinal sites anywhere in the body, the most important o' which is the liver, where the amoebae cause hepatic amoebiasis. 0-#*."&626 =$lac+ 'aeces @ upper (D2) gastrointestinal bleeding ($lood A!*l)7 =,ed 'aeces @ lower gastrointestinal bleeding (colon)7 =presence o' cold blood @ colorectal cancer, =rice wet stool @ cholera, =bloody, watery, mucoid stool @ amoeba (0tropho)oite @ in'ectious stage, 0cyst) %o detect the presence o' these parasites in humans, we undergo stool e<amination. %he steps are as 'ollows: 7ro## e:a"ination! 6pecimens should be e<amined: blood, mucus, intact worms or tapeworm segments. Sa"%le #i+e and %re%aration! 6pecimen si)e should be at least /g o' 'ormed 'eces (9/ cmH). 2' 'eces are so't, sample si)e should be Bg. 2' it is slurry-li+e, the sample should be Cg. -or li;uid 'eces, a sample o' KgA might be appropriate. 2nade;uate

sample si)e (e.g., 'ecal loop sample) may @ 'alse-negative. %o remove large 'ecal debris, sieving is recommended prior to centri'ugation. %he sample is sieved through cheesecloth or a tea strainer a'ter mi<ing with water or 'lotation solution. assive 'lotation +its typically include a device that prevents larger particles 'rom 'loating to the sur'ace. Flotation #olution! $oth the type and concentration o' sugar or salt solutions used can a''ect recovery o' diagnostic stages o' parasites 'rom 'eces. *ommon 'lotation solutes include sodium nitrate, )inc sul'ate, sucrose (usually granulated sugar), magnesium sul'ate, and sodium chloride. 6olutes can be mi<ed at varying concentrations with water to achieve 'lotation solutions with di''erent densities. -lotation solutions with higher densities are capable o' 'loating heavier (denser) parasite stages. !owever, higher density 'lotation solutions also 'loat many other 'ecal particles that can render preparations more di''icult to e<amine and can collapse thinshelled parasite stages, ma+ing them di''icult to identi'y or causing them to 'loat poorly. More viscous solutions, such as 6heather8s sugar (sucrose) solution, are more e''icient 'or centri'ugation. Most salt solutions dry very ;uic+ly, crystalli)ing on slides and obscuring observation. Centri,ugation! *entri'ugation o' sieved 'eces may be per'ormed in 'lotation solution either with a coverslip placed on top o' a 'illed tube or with the coverslip added a'ter the centri'uge has stopped. 2n the latter case, the tube is spun near-'ull, and then the tube is 'illed to 'orm a reverse meniscus, the coverslip is added, and the tube is allowed to sit a 'ew minutes longer. *entri'ugation with the coverslip on the tube wor+s best when a sugar 'lotation medium is used. .lternate methods 'or sampling the reverse meniscus include loops or glass rods that can be 'lamed between samples7 however, this approach is less e''icient than centri'uging with the coverslip in place. Slide e:a"ination! %he entire area under the coverslip should be e<amined. 2t is help'ul to 'ocus on a small air bubble to obtain the correct 'ocal plane. %he edge o' the coverslip can be sealed with nail polish to prevent drying and to allow e<amination o' the specimen under oil immersion. 6ucrose preparations can be stored in high humidity in a re'rigerator 'or hours to days without signi'icantly altering the morphology o' most common helminth eggs. .lthough routine 'ecal e<amination should always include centri'ugation, at times, other e<amination methods are needed to reach a diagnosis. -or e<ample, motile tropho)oites

and nematode larvae can be observed using the direct smear method. 4otes: 4ormal 6alt 6aline (466): used to 'ind the 'ollowing in 'ecal smears: o Negetable 'ibers o 4uclei 'ibers o 6tarch granules o -at globules "ugols 2odine: 3i''erentiate cysts o' amoeba *oncentration %echni;ues - used 'or 'inding microscopic structures such as parasite ova and cysts. (,emove 'at globules, starch granules, vegetable 'ibers, and nuclei 'ibers through sieving and centri'ugation.) 6ediments are to be observed under the microscope. Flotation" 'ind cysts and tropho)oites Sedimentations" 'ind the ova or di''erent parasites "aboratory testing o' 'eces: -eces sometimes re;uired 'or microbiological testing, loo+ing 'or intestinal pathogen, other parasite(disease. $iochemical tests done on 'eces include 'ecal elastase and 'ecal 'at measurements, as well as tests 'or 'ecal occult blood. 2t is recommended that the clinician correlate the symptoms and submit specimens according to laboratory guidelines to obtain results that are clinically signi'icant. -ormed stools o'ten do not give satis'actory results and suggest little o' actual pathological conditions. %hree main types o' microbiological tests are commonly done on 'eces: .ntibody-antigen type tests, that loo+ 'or a speci'ic virus (e.g. rotavirus). Microscopic e<amination 'or intestinal parasites and their ova (eggs). ,outine culture. (2nvolves strea+ing sample onto agar plates with special additives to inhibit the growth o' Dram-positive, thic+ membraned organisms and selectively allow enteric pathogens to grow, incubate them 'or a period, > observing the bacterial colonies that have grown. $ristol 6tool *hart: %he $ristol 6tool *hart Medical aid classi'y 'orm o' human 'eces into L categories. ,e'erred to as the PMeyers 6cale,P in the 51, developed

by 1.?. !eaton at the 5niversity o' $ristol and published in the Scandina)ian *ournal o+ Gastroenterology in /EEL. %he 'orm o' the stool depends on the time it spends in the colon. %he seven types o' stool are: /. 6eparate hard lumps, li+e nuts (hard to pass) B. 6ausage-shaped but lumpy H. "i+e a sausage but with crac+s on its sur'ace C. "i+e a sausage or sna+e, smooth and so't J. 6o't blobs with clear cut edges (passed easily) K. -lu''y pieces with ragged edges, a mushy stool L. ?atery stool, entirely li;uid. =/ > B @ constipation7 H > C @ Pideal stoolsP (especially C)7 J, K, > L @ diarrhea or urgency. *olor variations o' 'eces: Yello4: can be caused by an in'ection +nown as Giardiasis, which derives its name 'rom Giardia, an anaerobic 'lagellated proto)oan parasitetha t can cause severe and communicable yellow diarrhea. .nother cause o' yellowing is a condition +nown as Dilbert8s 6yndrome. %his condition is characteri)ed by Maundice and hyperbilirubinemia when too much bilirubin is present in the circulating blood. Blac?: resence o' ,$*s that has been in the intestines long enough to be bro+en down by digestive en)ymes. %his is +nown as melena, typically due to bleeding in the upper digestive tract, such as 'rom bleeding peptic ulcer. %he same color change (albeit harmless) can be observed a'ter consuming 'oods that contain substantial proportion o' animal bloods, such as $lac+ pudding or %i^t canh. *olor is caused by o<idation o' the iron in the blood8s hemoglobin. $lac+ 'eces can also be caused by a number o' medications, such as bismuth subsalicylate, and dietary iron supplements, or 'oods such as blac+ li;uorice, or blueberries. !ematoche)ia is similarly the passage o' 'eces that are bright red due to the presence o' undigested blood, either 'rom lower in the digestive tract, or 'rom a more active source in the upper digestive tract. .lcoholism can also provo+e abnormalities in the path o' blood throughout the body, including the passing o' red-blac+ stool. Blue: russian blue, used in the treatment o' radiation cesium and thallium poisoning, can turn the 'eces blue. .lso, substantial consumption o' products containing blue 'ood dye (things such as blue +oolaid or grape soda) Sil0er: . tarnished-silver or aluminum paintli+e stool color characteristically results when biliary obstruction o' any type (white

stool) combines with gastrointestinal bleeding 'rom any source (blac+ stool). .nd it can suggest a carcinoma o' the ampulla o' Nater, which will result in gastrointestinal bleeding and biliary obstruction, resulting in silver stool. -ecal contamination: . ;uic+ test 'or 'ecal contamination o' water sources or soil is a chec+ 'or the presence o' ($ coli bacteria per'ormed with the help o' Mac*on+ey agarplates or etri dishes. ($ coli bacteria uni;uely develop red colonies at temperature o' appro<imately CH V* (/FE V-) overnight. ?hile most strains o' ($ coli are harmless, their presence is indicative o' more serious 'ecal contamination, and hence a high possibility o' more dangerous organisms. -ecal contamination o' water sources @ highly prevalent, accounting 'or maMority o' unsa'e drin+ing water, which is the only water available to /./ billion people. 2n developing countries most sewage is discharged without treatment. #ven in developed countries events o' sanitary sewer over'low are not uncommon. %he main pathogens that are commonly loo+ed 'or in 'eces include: ,acteroides species Salmonella and Shigella -ersinia: incubated HFV* (IKV-), cooler than usual ampylobacter: incubated CBV* (/FIV-), special envi &eromonas andida: i' the person is immunosuppressed (undergoing cancer treatment) ($ coli O/JL i' blood is visible in the stool sample ryptosporidium (ntamoeba histolytica 032,#*% -#*." 6M#., (3-6)- test to chec+ an individuals stool 'or diseases /. lace small amount o' 'eces on a microscope slide. B. .dd drop o' li;uid to 'eces and mi< thoroughly. %ype o' li;uid added depends: =li;uid 'ecal sample 'or the presence o' proto)oan tropho)oites (live active proto)oa) @ saline (i' any e<tra li;uid is needed. =helminth eggs and proto)oan cysts in a small sample (bird droppings, rectal smear, etc) @ water or iodine. H. *over with a cover slip. Move the cover slip around until it lays 'lat. &ou should be able to read through the smear (light 'rom the microscope must be able to pass through the sample in order 'or you to e<amine it). C. #<amine the slide using the /FU obMective, and then go over it with the CFU obMective. $ecause this techni;ue e<amines only a very

small amount o' 'eces, it should only be used in the 'ollowing circumstances: a. "i;uid 'eces where proto)oan tropho)oites may be present. b. -ecal samples where amount o' 'eces obtained is too small to handle with any other techni;ue. c. .dMunct to a 'lotation techni;ue where you are loo+ing 'or eggs that do not 'loat. (May run ethyl acetate sedimentation and e<amine resultant pellet using the direct smear method.) arasitologist @! I""unologyASerology: MicrobiologyA!ematology7 .4& 24-#*%2O47 covers all aspects o' the immune system in all organisms. hysiological 'unction in states o' both health and disease. Mal'unction in immunological disorders (autoimmune diseases, hypersensitivities, immune de'iciency and transplant reMection.) hysical, chemical and physiological characteristics o' the components o' the immune system. #arly physicians characteri)ed organs later proven to be part o' the immune system be'ore the concept o' immunity (_immunis., "atin _e<empt) was developed. 1ey primary lymphoid organs o' the immune system: Y thymus and bone marrow 6econdary lymphatic tissues: Y spleen, tonsils, lymph vessels, lymph nodes, adenoids, and s+in =?hen health conditions warrant, immune system organs including the thymus, portions o' bone marrow and secondary lymphatic tissues can be surgically e<cised 'or e<amination while patients are still alive. =Many components o' the immune system@not associated with speci'ic organ7 cellular in nature are either embedded(circulating in various tissues located throughout the body. Serology: study o' blood serum to detect the presence o' antibodies against a microorganism. *ertain microorganisms stimulate the body to produce antibodies during an active in'ection. $lood is analy)ed in a laboratory to determine how certain antibodies react with speci'ic antigens. %he test can be used to con'irm the identity o' the speci'ic microorganism. 6everal serology techni;ues that can be used depending on the suspected antibodies: Yagglutination, precipitation, complement'i<ation, 'luorescent antibodies, and others. 6erology can determine e<posure to a particular microorganism7 not necessarily indicates a current in'ection. 4ormally, no antibodies are 'ound in the blood sample. .bnormal ,esults:

3etection o' antibodies: diagnose an active or previous in'ection or determine immunity to rein'ection by an organism. .s the diseaseYworse, antibodies. %ests may need to be repeated /F days to B wee+s a'ter i' a disease is suspected. 2' antibodies are 'ound, you may: 0 !ave a current in'ection 0 !ave been in'ected in the past 0 !ave immunity to an organism7 unli+ely to get sic+ 6ome o' the di''erent detectible diseases: 0 .moebiasis 0 .nthra< 0 $rucellosis 0 !uman immunode'iciency virus (!2N) 0 -ungal in'ection 0 Measles 0 ,ubella 0 ,6N 0 6yphilis 0 %ularemia 0 Niral hepatitis (various types) Other conditions under which test may be per'ormed: 0 .mebic liver abscess 0 -i'th disease 0 -ungal arthritis 0 Meningitis, cryptococcal 0 Meningitis, !. in'luen)a 0 Meningitis, meningococcal 0 Niral arthritis %ypes o' 2mmunity: a. 4atural 2mmunity: i. 4atural active: "ymphocytes activated by antigens on pathogen8s sur'ace7 when the person is e<posed to a live pathogen, develops the disease, and becomes immune as a result o' the primary immune response. ii. 4atural passive: Mother to child through the placenta or mil+7 during pregnancy, in which certain antibodies are passed 'rom the maternal into the 'etal bloodstream. b. .rti'icial immunity: 3egree and duration o' immunity depend on the type and amount o' antigen and how it enters the body. i. .rti'icial active: 2nMecting or oral inta+e. %a+es time 'or % and $ cells to be activated but gives long lasting immunity7 induced by a vaccine, a substance that contains the antigen. . vaccine stimulates a primary response against the antigen without causing symptoms o' the disease. ii. .rti'icial passive: 5sed during potentially 'atal diseases. rovides an instant response but only temporary as antibodies are not the body8s own so memory cells are not created. .n e<ample is tetanus inMection o' antito<ins given. .rti'icially ac;uired passive immunity is a short-term immuni)ation by the inMection o'

antibodies, such as gamma globulin, that are not produced by the recipient8s cells. 2mmune ,esponse o' the $ody to 2n'ection: *linical response may range 'rom nothing at all, through various degrees o' nonspeci'ic reactions, to speci'ic in'ectious disease. 2mmunologically, there is always a response, @ de'ense. 2' the de'ense is completely success'ul, there is no obvious bodily reaction7 i' it is partially success'ul, the a''ected person e<hibits symptoms but recovers 'rom an in'ectious disease7 i' unsuccess'ul, the person may be overwhelmed by the in'ectious process and die. %he immune system does this great Mob o' de'ending or +eeping people healthy and preventing in'ections. a. %he 2mmune 6ystem %he purpose o' the immune system is to +eep in'ectious microorganisms, such as certain bacteria, viruses, and 'ungi, out o' the body, and to destroy any in'ectious microorganisms that do invade the body. %he immune system is made up o' a comple< and vital networ+ o' cells and organs that protect the body 'rom in'ection. %he organs involved with the immune system are called the lymphoid organs, which a''ect growth, development, and the release o' lymphocytes - a type o' in'ection'ighting white blood cell. #ach lymphoid organ plays a role in the production and activation o' lymphocytes. "ymphoid organs include: adenoids appendi< blood vessels bone marrow lymph nodes lymphatic vessels eyer8s patches spleen thymus tonsils !ow lymphocytes 'ight in'ection: %he $ cells: produce speci'ic antibodies to speci'ic in'ectious microorganisms. %hese antibodies continue to e<ist in a person8s body, so i' the same antigen is presented to the immune system again, the antibodies are already there to do their Mob. #<ample: chic+enpo<. %he % cells: part o' the system that destroys antigens that have been tagged by antibodies or cells that have been in'ected or somehow changed. 1ill in'ectious microorganisms by +illing the body cells that are a''ected7 are involved in signalling other cells to do their Mobs: they release chemicals, called lympho+ines, which trigger an immune response to combat cancer or a virus. b. 2mmune response to Microorganisms

i. 2mmunity to $acteria: %he immune response to e<tracellular bacteria must counteract all o' the mechanisms o' invasion elicited by organisms7 includes antibodies to capsular polysaccharides, to e<oto<ins and to e<tracellular en)ymes7 .ntibodies to tetanus to<in or diphtheria to<in can neutrali)e the e''ects o' these to<ins and prevent host tissue destruction ii. 2mmunity to Niruses: .ntibodies capable o' binding directly to e<tracellular viruses may prevent viruses 'rom in'ecting other cells. 2' the virus has a viremic phase, neutrali)ing antibodies may be produced. .ntibodies can also diminish in'ectivity o' viruses by preventing attachment to the speci'ic receptor or by introducing con'ormational changes in the viral structure that promote aggregation iii. 2mmunity to -ungi: 2mmunity to 'ungi is primarily cell mediated. 3etection o' speci'ic 2gM and 2gD antibodies to certain 'ungi by immunoprecitin reactions can be help'ul in establishing the diagnosis and 'ollowing the course o' the disease. !owever, the antibodies do not play a protective role I""unoglo(ulin#: (2g) glycoprotein molecules produced by plasma cells in response to immunogen and 'unction as antibodies. /. Deneral -unctions o' 2mmunoglobulins a. .ntigen binding: bind speci'ically to one or a 'ew closely related antigens. #ach immunoglobulin actually binds to a speci'ic antigenic determinant. .ntigen binding by antibodies is the primary 'unction o' antibodies and can result in protection o' the host. b. #''ector -unctions: -re;uently the binding o' an antibody to an antigen has no direct biological e''ect. ,ather, the signi'icant biological e''ects are a conse;uence o' secondary Pe''ector 'unctionsP o' antibodies. %he immunoglobulins mediate a variety o' these e''ector 'unctions. 5sually the ability to carry out a particular e''ector 'unction re;uires that the antibody bind to its antigen. 4ot every immunoglobulin will mediate all e''ector 'unctions. 6uch e''ector 'unctions include: -i<ation o' complement $inding to various cells B. $asic 6tructure o' 2mmunoglobulins a. !eavy and "ight *hains: .ll immunoglobulins have a 'our chain structure as their basic unit. *omposed o' two identical light > two identical heavy chains. b. 3isul'ide bonds i. 2nter-chain disul'ide bonds - %he heavy and light chains and the two heavy chains are held together by inter-chain disul'ide

bonds and by non-covalent interactions %he number o' inter-chain disul'ide bonds varies among di''erent immunoglobulin molecules. ii. 2ntra-chain disul'ide binds - ?ithin each o' the polypeptide chains there are also intrachain disul'ide bonds. c. Nariable (N) and *onstant (*) ,egions ?hen the amino acid se;uences o' many di''erent heavy chains and light chains were compared, it became clear that both the heavy and light chain could be divided into two regions based on variability in the amino acid se;uences. %hese are: i. "ight *hain - N" (//F amino acids) and *" (//F amino acids) ii. !eavy *hain - N! (//F amino acids) and *! (HHF-CCF amino acids) d. !inge ,egion: %his is the region at which the arms o' the antibody molecule 'orms a &. 2t is called the hinge region because there is some 'le<ibility in the molecule at this point. e. 3omains: %hree dimensional images o' the immunoglobulin molecule show that it is not straight as depicted in 'igure B.. ,ather, it is 'olded into globular regions each o' which contains an intra-chain disul'ide bond ('igure B$-3). %hese regions are called domains. '. Oligosaccharides: *arbohydrates are attached to the *!B domain in most immunoglobulins. !owever, in some cases carbohydrates may also be attached at other locations.

H. 2mmunoglobulin -ragments: 6tructure(-unction ,elationships 2mmunoglobulin 'ragments produced by proteolytic digestion have proven very use'ul in elucidating structure('unction relationships in immunoglobulins. a. -ab 3igestion with papain brea+s the immunoglobulin molecule in the hinge region be'ore the !-! inter-chain disul'ide bond -igure C. ,esults in 'ormation o' two identical 'ragments that contain the light chain and the N! and *!/ domains o' heavy chain. .ntigen binding - %hese 'ragments were called the -ab 'ragments because they contained the antigen binding sites o' the antibody. #ach -ab 'ragment is monovalent whereas the original molecule was

divalent. %he combining site o' the antibody is created by both N! and N". .n antibody is able to bind a particular antigenic determinant because it has a particular combination o' N! and N". 3i''erent combinations o' a N! and N" result in antibodies that can bind a di''erent antigenic determinants. b. -c 3igestion with papain also produces a 'ragment that contains the remainder o' the two heavy chains each containing a *!B and *!H domain. %his 'ragment was called -c because it was easily crystalli)ed. c. -(ab8)B %reatment o' immunoglobulins with pepsin results in cleavage o' the heavy chain a'ter the !-! inter-chain disul'ide bonds resulting in a 'ragment that contains both antigen binding sites ('igure K). %his 'ragment was called -(ab8)B because it is divalent. %he -c region o' the molecule is digested into small peptides by pepsin. %he -(ab8)B binds antigen but it does not mediate the e''ector 'unctions o' antibodies. 0#"26.: #n)yme lin+ immunosorbent assay: rheumatoid arthritis, hepatitis, in'ections, mononucleosis, !2N(.236 0 *,: olymerase chain reaction-genes 2mmunologist B! Blood3(an?ing &I""uno3he"atology': 0$lood-typing 0*ross-matching 03onor 6election 2mmunohematology: branch o' hematology7 studies antigen-antibody reactions and analogous phenomena =relate to the pathogenesis and clinical mani'estations o' blood disorders. $e'ore ,$* trans'usion, blood samples 'rom donors and recipients are tested to determine their .$O group and 3 type, to detect une<pected ,$* alloantibodies, and con`rm cross-match compatibility. 2mmunohematology assays represent a subset o' pre-trans'usion testing, which also includes in'ectious disease serology, counting o' residual ?$*s in `ltered blood components, platelet cross-matching and additional assays. ,outine immunohematology testing has historically been per'ormed by centri'uging ,$*s and antibodies in test tubes 'ollowed by a visual determination o' the e<tent o' ,$* agglutination. .lthough standard tube testing is still considered by many to be the de 'acto [gold standard\ 'or immunohematology, this assay is labor intensive, not amenable to automation, and the results are operators dependent. ?ith a decline in laboratory technical sta'' trained to per'orm standard tube testing, this assay is becoming increasingly impractical. *urrent immunohematology testing methods have limitations including cost, throughput,

and adaptability to automation. -urthermore, current automated and semiautomated wor+stations cannot accommodate many other tests relevant to blood trans'usion. ABO Blood 7rou%# and 7rou%ing Sy#te": 3i''erences in human blood: due to presence( absence o' certain protein molecules called antigens and antibodies. %he antigens are located on the sur'ace o' the ,$*s and the antibodies are in blood plasma o' which individuals have di''erent types and combinations. %he blood group you belong to depends on what you have inherited 'rom your parents. %here are BFA genetically determined blood group systems +nown today, but the .$F and ,h systems are the most important ones used 'or blood trans'usions. 4ot all blood groups are compatible with each other. Mi<ing incompatible blood groups @ blood clumping or agglutination, which is dangerous 'or individuals. 4obel "aureate 1arl "andsteiner was involved in the discovery o' both the .$O and ,h blood groups. .ccording to the .$O blood group system there are 'our di''erent +inds o' blood groups: ., $, .$ > O: ,lood group &" . antigens on the sur'ace o' the red blood cells and $ antibodies in the blood plasma. ,lood group ,: $ antigens on the sur'ace o' the red blood cells and . antibodies in the blood plasma. ,lood group &,: both . and $ antigens on the sur'ace o' the red blood cells and no . or $ antibodies at all in the blood plasma. ,lood group /: neither . or $ antigens on the sur'ace o' the red blood cells but have both . and $ antibodies in the blood plasma. Rh Factor Blood 7rou%ing Sy#te": Many people also have a so called ,h 'actor on the red blood cell8s sur'ace. %his is also an antigen and those who have it are called ,hA. %hose who do not have it are called ,h-. . person with ,h- blood does not have ,h antibodies naturally in the blood plasma (as one can have . or $ antibodies, 'or instance). $ut a person with ,h-blood can de)elop ,h antibodies in the blood plasma i' he or she receives blood 'rom a person with ,hA blood, whose ,h antigens can trigger the production o' ,h antibodies. . person with ,hA blood can receive blood 'rom a person with ,h- blood without any problems. Blood 7rou% Notation#: . ,hA . ,h$ ,hA $ ,h.$ ,hA .$ ,hO ,hA O ,h-

-i,,erent (lood co"%onent# and ho4 they are u#ed: 4ormally, LS-IS o' human body weight is 'rom blood. *arries out critical 'unctions o' transporting OB and nutrients to our cells and getting rid o' *OB, 4!H, and other waste products. lays a vital role in our immune system and in maintaining a relatively constant body temperature. $lood is a highly speciali)ed tissue composed o' more than C,FFF di''erent +inds o' components. -our o' the most important ones are red cells, white cells, platelets, and plasma. .ll humans produce these blood components: no populational or regional di''erences. ,ed blood cells or erythrocytes: relatively large microscopic cells without nuclei7 similar to the primitive pro+aryotic cells o' bacteria. CFS-JFS o' the total blood volume. %hey transport OB 'rom the lungs to all living tissues o' the body and carry away *OB. roduced continuously in our bone marrow 'rom stem cells at a rate o' about B-H million cells per second. 6tem cell: embryonic cells that have not yet become speciali)ed tissue cells--they potentially can develop into any type o' tissue in the body. *hildren and adults retain somewhat speciali)ed stem cells in their bone marrow. %hese stem cells are the source o' the maMor blood cells-erythrocytes, leu+ocytes, and thrombocytes (platelets). !emoglobin: gas transporting protein molecule that ma+es up EJS o' a red cell. #ach red cell has about BLF,FFF,FFF iron-rich hemoglobin molecules. eople who are anemic generally have a de'iciency in red cells, and subse;uently 'eel 'atigued due to a shortage o' o<ygen. !uman 'etal hemoglobin molecules di''er 'rom those produced by adults in the number o' amino acid chains. -etal hemoglobin has three chains, while adults produce only two. .s a conse;uence, 'etal hemoglobin molecules attract and transport relatively more o<ygen to the cells o' the body. ?hite cells or leu+ocytes: e<ist in variable numbers and types but ma+e up a very small part o' blood8s volume- 9/S in healthy people. 4ot limited to blood, they occur elsewhere in the body as well, most notably in the spleen, liver, and lymph glands. Most are produced in bone marrow 'rom the same +ind o' stem cells that produce red blood cells. Others are produced in the thymus gland, which is at the base o' the nec+. "ymphocytes are the 'irst responders 'or our immune system. -%hey also have the 'unction o' getting rid o' dead or dying blood cells as well as 'oreign matter such as dust and asbestos. ,ed cells remain viable 'or only about C months be'ore they are removed

'rom the blood and their components recycled in the spleen. %here are 'ive important types o' white blood cells: ,asophils" release histamines$ (osinophils [eat\ other cells. %he technical term 'or the eating o' a cell is phagocytosis, so eosinophils are said to phagocyti)e comple<es 'ormed between antigens (the invading o''ender) and antibodies (a [home team\ de'ender). Lymphocytes" +ill cells with viruses. "ymphocytes scan the body loo+ing 'or viruses. %here are two types o' lymphocytes: $ cells and % cells. % cells are the type o' virus hunters measured in a person with ac;uired immunode'iciency syndrome (.236). Monocytes" precursors to macrophages, [big eater.\ Macrophages digest bacteria and viruses. 'eutrophils: most abundant white blood cells in the body. %hese cells phagocyti)e bacteria, and in doing so +eep your system 'rom being overrun by every germ with which it comes into contact. latelets or thrombocytes: cell 'ragments without nuclei that wor+ with blood clotting chemicals at the site o' wounds. .dheres to the walls o' blood vessels, and plugging the rupture in the vascular wall. *an release coagulating chemicals which cause clots to 'orm in the blood that can plug up narrowed blood vessels7 help 'ight in'ections (release proteins that +ill invading bacteria and other microorganisms.) latelets stimulate immune system. 9a si)e o' ,$* lasma: relatively clear, yellow tinted water (EBAS), sugar, 'at, protein and salt solution which carries the red cells, white cells, and platelets. 4ormally, JJS o' our blood8s volume is made up o' plasma. .s the heart pumps blood to cells throughout the body, plasma brings nourishment to them and removes the waste products o' metabolism. lasma also contains blood clotting 'actors, sugars, lipids, vitamins, minerals, hormones, en)ymes, antibodies, and other proteins. %wo maMor proteins contained in plasma are" Damma globulin (immunoglobulin): broad term 'or a class o' proteins that ma+e up the di''erent types o' antibodies. %he production o' antibodies, (help 'ight in'ections), is controlled by immune system. -ibrinogen: protein involved in blood clotting. I"%ortant %rocedure# done related in the I""unohe"atology or (lood (an?ing #ection:

%iming o' *ompatibility %esting: . sample must be obtained 'rom the patient within H days o' the scheduled trans'usion 'or compatibility testing i' any o' the 'ollowing conditions e<ist: atient has been trans'used with a blood component containing red blood cells in the preceding H months atient pregnant within the preceding H months atient history is uncertain %esting o' a new sample is necessary because a patient can develop a primary antibody response at any time within the 'irst three months 'ollowing immuni)ation. .$O %yping: accomplished by: %esting patient8s red cells with anti-. and anti-$ antisera ('orward typing) %esting patient8s serum 'or anti-. and anti-$ (bac+( reverse typing) %he .$O system is uni;ue because plasma has naturally occurring antibodies to the .$O red cell antigens that are absent 'rom his or her own red cells. .ntibodies are the basis 'or .$O compatibility criteria when selecting red cells and plasma 'or trans'usion. Inter%retation o, ABO ty%ing RBC Seru Seru ABO Co"%ati C "C "CB 7rou% (le RBC# Anti3 A cell# B cell# A . .,O A A $ $,O A .$ .$,.,$,O A A O Only O 3etermination o' .$O blood groups @ most important pre-trans'usion compatibility test. 2' tests are done to insure that donor and recipient belongs to the same .$O blood group, even i' no other tests are done, the donor8s red blood cells will be compatible with the recipient8s plasma in about ELS o' cases. 6election o' ,$* 5nits ?hen .$O 6peci'ic $lood is not available: 2mportant to use ,$*s that are compatible with the recipient8s serum7 crossmatch compatible. 3onor ,$*s must not contain . or $ antigens that react with the anti-. or anti-$ present in the recipient8s serum. 2n this situation the large amount o' anti-. or anti-$ in the recipient8s plasma would bind to trans'used .$O incompatible ,$*s and cause hemolysis. %he reverse situation: recipient8s ,$*s have antigen that reacts with an antibody in the donor8s plasma @ not as important. %he small amount o' antibody present in the /FF m" o' plasma remaining in a ,$* unit is rapidly diluted about HFU in the recipient8s plasma be'ore the antibody can inMure enough ,$*s to be clinically apparent. RBC #election 4hen ty%e3#%eci,ic (lood i# una0aila(le #t Reci%i Cho 1nd Cho 8rd Cho ent ice ice ice

ABO 7rou% OA .A .$A Reci%i ent ABO 7rou% .$-

RBC $nit OA .A .$A #t Cho ice RBC $nit .$-

RBC unit OOA .A 1nd Cho ice RBC unit OO-

RBC unit 4one O-

$A 8rd Cho ice RBC unit .A $A or OA .$.$.$$lood *rossmatch: ($*M) is per'ormed to detect serological incompatibility by identi'ying antibodies in donor or recipient plasma against recipient or donor red blood cells. . $*M is divided into two parts: the maMor crossmatch consists o' mi<ing the patients plasma with the donors red blood cells7 the minor crossmatch consists o' mi<ing the donors plasma with the patients red blood cells. O' the two tests, the maMor blood crossmatch is much more important in determining survival o' the trans'used red blood cells. rocedure: /. *ollect blood into an #3%. tube 'rom recipient and potential donor(s). *entri'uge (/FFF < g 'or J min) to separate plasma 'rom ,$*s, remove add CGJ ml o' $6, mi< well, centri'uge /GB minutes, remove saline, leaving pellet o' ,$*s at bottom o' tube. B. %a+e plasma 'rom each sample with a pipette and trans'er plasma to clean, labeled glass or plastic tubes. 4ote any hemolysis. H. ?ash ,$*s H times with $6: C. ,esuspend ,$* pellet with $6 to ma+e a HGJS ,$* suspension. J. repare 'or each donor H tubes labeled maMor, minor, and recipient control. .dd to each tube B drops (JF bl) o' plasma and / drop (BJ bl) o' ,$* suspension as 'ollows: a. maMor recipient plasmaAdonor ,$*s, b. minor donor plasmaArecipient ,$*s, c.recipient control recipient plasmaArecipient ,$*s. K. Mi< gently and incubate 'or /JGBF minutes at HLV * in a warm water bath. L. *entri'uge 'or /J seconds at /FFF < g. I. #<amine supernatant 'or hemolysis. E. Dently resuspend the button o' ,$*s by tapping tube and e<amine 'or macroscopic agglutination. *lassi'y as /A ('ine), BA (small), HA (large), or CA (one large agglutinate). .n autocontrol sample o' recipient ,$*s and plasma is included because some recipients may have autoagglutination inter'ering with

the $*M. 2' the recipient control is positive (i.e., agglutination is present), one cannot draw conclusions about blood compatibility between patient and donors. .ny hemolysis and(or agglutination in the maMor or minor $*M (but not the control) indicate an incompatibility and the need to choose a new donor. . compatible $*M does not prevent sensiti)ation or delayed trans'usion reactions7 it simply indicates that at the present time no detectable antibodies against the ,$*s e<ist. 1 *ind# o, Cro## Matching MaDor Cro##"atch - this is the most important crossmatch, comparing donor erythrocytes to recipient serum (i.e. you are chec+ing 'or pre'ormed (ac;uired or naturally occurring) antibodies in recipient serum against donor erythrocytes. -or the maMor crossmatch, you need red blood cells 'rom the donor (this can be whole blood 'rom a donor animal or pac+ed red blood cells) in #3%. or citrate and serum 'rom the recipient (non-anticoagulant tube). Minor Cro##"atch- %his compares donor serum to recipient erythrocytes and chec+s 'or pre'ormed antibodies in donor serum that could hemolyse recipient red cells. %his crossmatch is less important as usually the donor serum is mar+edly diluted a'ter trans'usion and is unli+ely to produce a signi'icant trans'usion reaction. %his type o' crossmatch could be important i' trans'using small patients, in which hemodilution is less li+ely to occur. 3onor 6election: procedure established to evaluate and assess the health status and ris+ 'actors o' the potential donors o' biological material li+e blood. 3onors are selected: health will not be compromised A donated materials are sa'e 'or reuse in the recipients. *are'ul donor selection plays a maMor role in determining whether a unit will be e''ective and 'ree o' transmittable disease or not. 2n 3onor 6election, a limited physical e<amination A detailed medical history must be done on the day o' and be'ore each donation. %he se;uence in which e<aminations are done or ;uestions are as+ed may be arranged 'or convenience. %he interviewer must be ;uali'ied personnel, as designated or assigned by the medical doctor, in a manner that assures auditory and visual privacy, relieve an<iety, and allows time 'or any necessary e<planation. .nswers must be recorded as [&es\ or [4o\ with some details as indicated. ,esults o' the e<aminations must also be recorded. ReEuire"ent# and 2rocedure# in -onor Selection Weight" 3onors weighing //Flbs (JF+g) or more may give CJFml A(- CJ ml o' blood A up to HF ml 'or processing tubes. 3onors QJF+g not normally drawn.

 !ulse" %he pulse should be counted at least /J seconds. 2t should be between JF and /FF beats per minute. 2t should not e<hibit any pathologic irregularity. !owever, i' a potential donor is an athlete with high e<ercise tolerance, a lower pulse rate may be acceptable. %he blood ban+ physician should evaluate mar+ed abnormalities o' pulse and recommend acceptance, reMection, or re'erral 'or additional evaluation. ,lood pressure: %he systolic blood pressure should not be higher than /IF mm !g and the diastolic blood pressure should be no higher than /FF mm !g. Skin lesions" %he s+in at the site o' venipuncture must be 'ree o' lesions. $oth arms must be e<amined 'or signs o' drug abuse. *ommon 'indings would be needle puncture mar+s and(or sclerotic veins. Mild s+in disorders such as acne or rash not necessarily causes o' de'erment unless present in the antecubital area ( unusually e<tensive. 3onors with boils, purulent wounds, or severe s+in in'ections anywhere on the body should be de'erred. 2n addition, those with purplish-red or hemorrhagic nodules or indurated pla;ues suggestive o' 1aposi8s sarcoma (a cancer that develops 'rom the cells that line lymph or blood vessels). General appearance" 2' the donor loo+s ill, appears to be under the in'luence o' drugs and(or alcohol, or is e<cessively nervous, best to de'er temporarily. #emperature" %he oral temperature must not e<ceed HL.Jo*. 5se caution when ta+ing the patient8s temperature with a glass thermometer. 5sing a thermometer cover is advised in doing so. 0ematocrit or 0emoglobin" %hese values vary depending on the source. ,e'er to the table below: 6ource o' %est Male -emal 6pecimen Method e #arlobe !gb /H g(dl /B.J !ct HES g(dl HIS -inger or vein !gb /B.J /B.F !ct g(dl g(dl HIS HKS opper sul+ate method o+ screening" place a drop o' blood into a test tube o' copper sul'ate solution7 i' the drop o' blood sin+s to the bottom in an acceptable amount o' time, the donor ;uali'ies. 2' the drop o' blood 'loats or ta+es too long to sin+ the donor is de'erred. %est procedures to be used are adapted to the source o' the specimen. %he solutions should be stored at room temperature in tightly capped containers to prevent evaporation. -or routine use, dispense HF ml solution into a labeled, clean, dry tube or bottle. *hange solution daily >

a'ter every BJ tests. ,esults that indicate satis'actory hemoglobin levels are usually accurate7 some results that indicate hemoglobin levels are 'alse. !rocedure +or Finger stick" -inger stic+ method is a procedure in which a 'inger is pric+ed with a lancet to obtain a small ;uantity o' capillary blood 'or testing. %his also called a 'inger pric+. 6teps 'or the method: /) *lean the site o' s+in puncture thoroughly with antiseptic solution and wipe with sterile gau)e. B) uncture a 'inger slightly to the side with sterile disposable lancet. (Dood 'ree 'low o' bloodc) H) 3o not s;uee)e the 'inger (dilutes the drop with e<cess tissue plasma and give 'alse results.) C) *ollect blood in a capillary tube (dont allow airc) J) "et one drop o' blood 'all gently 'rom the tube at a height o' about / cm above the sur'ace o' the proper sul'ate solution. Observe 'or /J seconds. ,ecord results less than /B.J gm or greater than /H.J gm, and so 'orth. Tran#,u#ion Reaction#: &cute 0emolytic Reactions 1234 0ours5: 2ncompatible blood administration results in an antigen(antibody response with activation o' complement and subse;uent intravascular hemolysis. Sepsis6,acterial ontamination" %rans'usion o' bacterially contaminated blood components. #rans+usion-Related &cute Lung 7n8ury 1#R&L75" *ommonly results 'rom the in'usion o' donor antibodies directed against recipient !". class 2 or 22 antigens or neutrophil antigens. %he antigen( antibody comple< activates complement with resultant neutrophil in'lu< into the lungs. 4eutrophil activation causes capillary lea+age and pulmonary damage. 2n're;uently, recipient antibodies against cognate donor antigens may be implicated. &llergic 1Se)ere5 &naphylactic or &naphylactoid Reactions" %he trans'usion recipient has an antibody which may be an 2g# directed against an antigen in donor plasma, such as an 2g.-de'icient patient who possesses antibodies to 2g.. %he cause, however, is o'ten not identi'ied. #rans+usion-&ssociated irculatory /)erload 1#& /5" a li'e-threatening condition7 rapid increases in blood volume in patients with compromised cardiac or pulmonary 'unction and(or in patients with chronic anemia and e<panded plasma volumes. Febrile 'onhemolytic Reactions" re'ormed anti-!". antibodies in the recipient ('rom

pregnancy or previous trans'usion) react with corresponding antigens on trans'used white blood cells or platelets and trigger cyto+ine release. .lternatively, pre'ormed cyto+ines 'rom white blood cell brea+down in the donor units may be directly in'used. Most 'ebrile nonhemolytic reactions are benign7 some cause signi'icant discom'ort > hemodynamic ( respiratory changes. &llergic 1Mild5 Reactions: %he trans'usion recipient usually has an 2g# antibody on mast cells directed against an antigen in donor plasma resulting in activation and release o' histamine. Gra+t-)s-0ost 9isease 1G:095: can occur i' !". incompatible with the trans'usion recipient, mount an attac+ against the recipients tissues, causing enterocolitis, rash, and pancytopenia. 3elayed !emolytic ,eactions (OBC !ours): patient had antibody against a red cell antigen in the past. %he titer o' this antibody has decreased to below detectable levels7 so antibody screen per'ormed prior to trans'usion didnt detect antibody. !osttrans+usion !urpura 1!#!5: %hrombocytopenia occurs in a patient who has made an antibody against a 'oreign platelet antigen as a result o' pregnancy or a previous trans'usion. .'ter a trans'usion o' red cells or platelets, antibodies attach to sur'ace antigen sites on platelets, resulting in their destruction by splenic and liver macrophages. 2mmunohematologist- blood typing, crossmatching, prepares blood components. athologist (M3 clinical > anatomic pathology) Y Medical %echnologist or *linical "aboratory 6pecialist Y Medical %echnician athologist: study the cause and development o' disease Medical %echnologist: e<ercises technical and scienti'ic 'unctions in medical laboratories Medical %echnician: in charge o' operating speciali)ed tools and e;uipment and administering tests on patients hlebotomist: ta+e blood samples 'rom patients 'or testing in laboratories. *ytotechnologist: study cells and cellular anomalies. !istotechnologist: prepares tissue specimens o' human and animal origin 'or a pathologist to e<amine *ytogenetic technologist: study normal and abnormal chromosomes in cells, and their relationship to disease. ME-ICAL TECHNOLO7Y *linical "aboratory 6cience. 5se o' technology to evaluate health status.

.u<iliary branch o' medical science because

o' diagnostic relevance Y stepping stone to medical career. ".$O,.%O,&, !#."%!, 32.D4O626, %,#.%M#4%. 2denti'y the nature o' conditions to increase li'espan and improve the ;uality o' li'e. .pplication o' natural, physical and biological sciences to the per'ormance o' lab procedures. er'ormance o' lab determinations and analyses to diagnose and treat disease and to maintain health. Obtaining necessary in'ormation. .u<iliary branch o' laboratory medicine which deals with e<amination og chemical, microscopic, bacteriologic and other medical laboratory procedures or techni;ues 'or a physician to diagnose, study, and treat disease as well as promote health. N."5# O- "2-#.

Highlights in the History of Medical Technology .. Early Beginning o, Medical Technology /. E0idence# ,ro" Re#earche# (y Noted MedTech# 5i0ian Herric? G (/JFF $*) arasites: #aenia ; &scaris$ [#bers apyrus\ boo+ 'or the treatment o' diseases, contains three stages o' hoo+worm in'ection. Ruth 6illia"# F & GH>3 ;8B' An Introduction to the Profession of Medical Technology, believes that the science began during Medieval eriod. 5rinalysis was a 'ad. 2t was the oldest laboratory procedure$ (3iabetes noted as KFF $*. #arly !indu doctors noticed ants attracted to urine.) Anne Fagel#on G (/Cth century) 2talian physician at the 5niversity o' $ologna employed Ale##andra 7iliani! 3ied o' laboratory ac;uired in'ection. 1! Noted Scienti#t# in the Early -e0elo%"ent o, MedTech Hi%%ocrate# F -ather o' Medicine. -ormulated !ippocratic Oath- code o' ethics 'or physicians. -our [humors\ or body 'luids in man: blood, phlegm, yellow bile > blac+ bile. G serve as the source o' a persons disposition and disease in the ancient times. !e mentioned tuberculosis, malaria, mumps, anthra< and purpura septicaemia (childbed 'ever) in his writings. 2o%e Innocent 5II G 'irst recipient o' blood draught. (d) 6illia" Har0ey G discovered blood circulation. %he era o' blood trans'usion started. Richard Lo4er G animal to animal blood trans'usion.

 Iean Ba%ti#te3-eni# G animal to human Ia"e# Blundell G human to human

trans'usion. *arl Land#teiner G discovered .$O blood group system. Anton 5an Leeu4enhoe? G invented compound microscope. -irst to describe red blood cells, to see proto)oa and classi'y bacteria according to shape. Marcello Mal%ighi G greatest o' all early microscopists. !is wor+ in embryology and anatomy de'ined him [Founder o+ !athology.\ Rudol, 5ircho4 G practice o' pathology is only in his time. One o' the youngest medical specialists. !e 'ounded the &rchi)es o+ !athology in ,erlin in <=4>! Her"an Fehling G per'ormed the 'irst ;uantitative test 'or urine sugar. Iule# -u(o#eE G in <=?4 he developed the 'irst vi#ual colori"eter based on $eers "aw (or $eer-"amberts "aw, one o' the guiding principles in most clinical chemistry assays nowadays: [#he concentration o+ substance is directly proportional to the amount o+ light absorbed or in)ersely proportional to the logarithm o+ the transmitted light,\) but was introduced in /EFB. .Aniline dye# F Made staining and microscopic study on bacteria possible. %his led to the development o' microbiology Loui# 2a#teur F asteuri)ation! Ro(ert *och F %$ and *holera Ale:ander Fle"ing F 2enicillin $. The Hi#tory o, MedTech &$NITE- STATES' -r! Sila# H! -ougla# F established 'irst chemical laboratory in the 5niversity o' Michigan in /ICC. Carl 0on 5oit F 'irst hygienic laboratory in Munich! -r! 6illia" H! 6elch F B@B9 established another laboratory in $ellevue !ospital Medical *ollege. 2t consisted o' three rooms with +itchen tables and si< anti;ue microscopes. !e gave the +irst Laboratory course in an &merican medical school$ <88 , he became the 'irst pro'essor o' pathology at Zohn !op+ins 5niversity where he taught pathology, bacteriology and e%perimental pathology 1autopsy!" Her"ann N! Bigg# and 6illia" H! 2ar? G headed, in BH19 the 'irst public diagnostic bacteriologic laboratory established by the NYC -e%art"ent o, HealthJ the La(oratory o, Hygiene at the 5niversity o' ennsylvania was opened. -r! 6illia" O#ler F opened the First linical Laboratory in <=@A at *ohn 0opkins 0ospital! 6pecial attention being given to the search 'or Malarial !arasites in the blood. The 6illia" trans'usion.

2e%%er la(oratory also opened in 5niversity o' ennsylvania. -r! Si"on Fle:ner G -irst 2athologi#t in o, Iohn Ho%?in# Ho#%ital -e%art"ent o, 2athology! HGG cen#u#3 /FF technicians9 all "ale9 4ere e"%loyed in the $S! Increa#ed to 89<GG in H1G 4here >GK 4ere ,e"ale#! T4o year# later9 89G8< ho#%ital# had clinical la(oratorie#! HGB cen#u# G 3r. Zames *. %odd wrote [. manual o, Clinical -iagno#i#L! Retitled MClinical -iagno#i# (y La(oratory Method#L in it# >th edition (y -r! Todd and -r! Arthur San,ord! H G 2nsurance act was approved. H < G State o, 2enn#yl0ania reEuired all ho#%ital# and in#titution# to ha0e an adeEuate la(oratory and ,ull3ti"e la(oratory technician! H B G The A"erican College o, Surgeon# conducted the ,ir#t in#%ection# o, ho#%ital! Out o, >@ only B %a##ed initially and another GG on rein#%ection! Iohn *ol"er G published [%he 3emand 'or and training o' "aboratory %echnicians\ this includes the 'irst 'ormal training course in medical technology. Fir#t call ,or a "ethod o, certi,ying technologi#t# on a national #cale! H1G G *linical la(oratorie# (eca"e di#tinct ad"ini#trati0e unit# o, #er0ice directed (y a chie, %hy#ician! It con#i#t o, ;A< #ection#: #ioche$istry, clinical %athology, #acteriology, serology&i$$unology and radiology! H1 G 3enver 6ociety o' *linical athologists was organi)ed. H11 G . course bulletin entitled ['ourses in Medical Technology for 'linical and (a#oratory TechniciansL 4a# i##ued! H18 G 5niversity o' Minnesota was the 'irst to o''er a degree level program. .merican 6ociety o' *linical athology (.6* ) was 'ounded. H1B G .6* established its $oard o' ,egistry. H81 G .merican 6ociety o' *linical laboratory %echnicians, precursor o' .merican 6ociety o' Medical technology was 'ounded. H8; G hysicians hired technical assistants to help in their wor+. H8< G .6* $oard o' ,egistry re;uired a college degree 'or a medical technology certi'ication. H;G G 56 re;uired two3year collegiate education and 13"onth training! H<G G standard curriculum was 'ormali)ed in preparation 'or a $achelor o' 6cience degree. 66II G & H8H3 H;<' Mclo#ed #y#te" L o, (lood collection 4a# 4idely ado%ted!

 H>@ G *linical "aboratory 2mprovement .ct o'

/EKL! Mini"u" Euality reEuire"ent#9 regulation! National Co""ittee ,or Clinical La(oratory Standard# &NCCLS' G a grou% o, clinician# and la(oratory #cienti#t# to di#cu## 4ay# o, i"%ro0ing %atient #er0ice#! H@< G M%s are re;uired to have $achelor 3egree or the e;uivalent. 2' not the will ta+e pro'iciency e:a" (y HE6! ReEuire"ent# 4ere High #chool di%lo"a and ,our year# o, la(oratory e:%erience! The te#t include#: he$atology, #lood #an)ing, clinical che$istry and $icro#iology! H@@ G .merican 4ational 6tandards 2nstitute accredited 4**"6 and became the home o' 4ational Re,erence Sy#te" ,or the Clinical La(oratory! HH1 G *linical "aboratory 2mprovement .mendments o' /EII were implemented. .ll laboratories are re;uired to have certi'ication (y the -e%art"ent o, Health and Hu"an Ser0ice#! *. The Hi#tory o, MedTech &2HILI22INES' #stablished at the end o' ??22, by the BKth Medical "aboratory o' the 56 .rmy. -irst clinical laboratory at ]uiricada street, 6ta. *ru), Manila. ?here the current Manila ublic !ealth "aboratory is located. 3o! rendered lab non-'unctional 'or some time a'ter .mericans le't. -r! 2io de Roda (bacteriologist) 'ormally organi)ed the M !" 'rom the remnants o' old lab in October /, /ECJ. .ssisted by -r! Mariano Ica#iano who was the Manila *ity !ealth O''icer. H;@, the high school graduate training was revived under 3r. io de ,oda and 3r. rudencia 6ta. .na, but trainees were unmotivated. H<;, si< month lab training with certi'icate. 3r. 6ta. .na prepared syllabus, 3r. %irso $riones Moined the two doctors in the training program. 323 4O% ".6% "O4D. $6 Med%ech (J years) was o''ered by the !hilippine Bnion ollege 1&d)entist Bni)ersity o+ the !hilippines5 and Manila Sanitarium 1Manila &d)entist Medical enter$5 ioneering e''ort o' Mr#! 6illa Hilgert Hedric?, missionary o' 6eventh 3ay .dventist *hurch in the hilippines. CFounder o+ Medical #echnolody (ducation in the !hilippines$. .pproved by the $ureau o' #ducation (3#*6) in /EJC. 5*s 'irst graduate in /EJK, -r! Ie##e $"aliNsuccess'ul O$-gynecologist and owner o' the Omega "aboratory at Nito *ru). /EJL-/EJI G -r! Antonio 7a(riel and -r! 7u#ta0o Reye# o''ered M% as an #lective 'or Cth > Jth year $6 harma students in $ST. ,ev. Fr! Loren+o Rodrigue+ decided to o''er is as a course. Zune /L, /EJL, a temporary permit

was issued by the 3ep#d 'or /st to Hrd year students and in Zune /EKF, the permit 'or the internship program was issued. -ull recognition o' the C-year $6 Med%ech was given on Zune /C, /EK/. CE$ 'ollowed in /EKF through the e''orts o' Mr#! 2uri,icacion Sunico3Suaco, who was granted by the 5niversity resident Car"en de Luna, to wor+ on the 'easibility o' o''ering Med%ech. 5pon the approval o' the $ureau o' #ducation, Mrs. 6uaco became the dean 'rom /EKF-/EKH. -irst graduates in /EKB. FE$ started Med%ech in /EK/ through -r! Horacio A Ylagan and -r! Sera,in I! Iuliano through the authority granted by the late 3r. "auro !. anganibana nd 3r. Zesus $. 4olasco, 3ean and 6ecretary o' the 2nstitute o' Medicine respectively. -ormally opened when the $ureau o' #ducation approved on Zuly J, /EKB. 3r. &lagan automatically became %echnical 3irector o' the school. -irst graduates in /EKH. *urrently appro<imately KF collegesc 5 o''ers the similar course, BS 2u(lic Health! ostgraduate studies: The 7raduate School and the 2hili%%ine 6o"enO# $ni0er#ity o''er MS in MedTech. $2 o''ers /-year, nonthesis Ma#ter in 2u(lic Health. 0(d emphasiDes need o+ post-grad study in tertiary +aculty members. 2t is reviewed in accreditation i' a school o' Med%ech wants to upgrade its status.

EM2LOYMENT O22ORT$NITIES OF ME-TECH 7RA-$ATES: Dovt and private hospitals, clinical laboratories and blood ban+s. 6ales and industry: sales representatives, , representatives or educational representatives 'or their company, or as part o' a health program 'or employees. ,esearch: industrial (new products and testing prior to distribution), medical center research may involve development or education 'or new laboratory methods, new clinical treatment method and varying types o' investigation. #ducation: !igh school or college in *hemistry, Mathematics, or $iological 6ciences or Medical 6ciences. Neterinary medicine: opportunities in research and in a veterinarians o''ice. .broad: 56. and the Middle #ast. 6tepping stone 'or Medicine. 2ERSONAL TRAITS OF A ME-ICAL TECHNOLO7IST: 6ervice oriented. atience. !onesty. .ccuracy.

 6+ills. 3edication. #motional Maturity.

U -actor. CO-E OF ETHICS OF 2AMET: &Re0i#ed9 HH@9 Chang' .s 2 enter the practice o' Medical %echnology, 2 shall: .ccept the responsibilities inherent to being a pro'essional 5phold the law and shall not participate in illegal wor+ .ct in a strict spirit o' 'airness to all and in a spirit o' brotherhood towards other members o' the pro'ession .ccept employment 'rom more than one employer only when there is no con'lict o' interest er'orm my tas+ with 'ull con'idence, absolute reliability and accuracy 6hare my +nowledge and e<pertise with my colleagues *ontribute to the advancement o' the pro'essional organi)ation and other allied health organi)ations ,estrict my praises, criticisms, views and opinions within constructive limits %reat any in'ormation 2 ac;uire in the course o' my wor+ as strictly con'idential 5phold the dignity and respect o' my pro'ession and conduct mysel' a reputation o' reliability, honesty, and integrity $e dedicated to the use o' clinical laboratory science to promote li'e and bene'it man+ind. ,eport any violations o' the above principles o' the pro'essional conduct to authori)ed agency and to the ethics committee o' the organi)ation %o these principles, 2 hereby subscribe and pledge to conduct mysel' at all times in a manner be'itting the dignity o' my pro'ession. Acco"%li#h"ent# o, 2ASMETH: * # (*ontinuing ro'essional #ducation) rogram 'or Medical %echnology 'aculty. reparation o' 6tandard *urriculum 'or $6 M% 6chools. reparation o' 6tandard *ourse syllabus 'or pro'essional subMects in M%. 6cholarship grants 'or M% students. *ommunity outreach program. ,ecognition to $6 M% *ourse: .6M#%! gold medal 'or e<cellence award. .ccreditation as * # rovider 'or ,M%s. 6trong association o' schools through annual .6M#%!- .M#% ;ui) shows.