IMPLANT TRAINING

IN
DEPARTMENT OF LABORATORY SERVICES

Meenakshi Mission Hospital & Research Centre

Lake area, Melur road, Madurai -625 107.
Report submitted in partial fulfillment for the degree of

BACHELOR OF TECHNOLOGY
IN
BIOTECHNOLOGY

Submitted by
G. ABISHA (9916001006) T.TVARETA (9916001137)

F. ANGELIN JENIT (9916001011) N.S. SUPRAJA (9916001155)

R. MARI SELVA SUNDARI S. MARIA AGNES ROGANZIA

(9916001068) (9916001069)

DEPARTMENT OF BIOTECHNOLOGY
Kalasalingam Academy of Research and Education)
Accredited with ‘A’ Grade with NAAC
ANAND Nagar, KRISHNANKOIL-626126.
1
 CERTIFICATE
This is to certify that the report submitted during the IMPLANT TRAINING in
partial fulfillment of the requirements for the award of the degree of “B.TECH
IN BIOTECHNOLOGY” is a record carried out by the students under my
supervision and guidance and that no part of the report has been submitted for the
award of any degree, diploma or fellowship or other similar titles.

Place: Madurai

Date:

Head of the Department External Examiner

2
 DECLARATION

We hereby declare that the work embodied in the dissertation has been originally
carried out by us, under the guidance and supervision of Mr.
Muthuchelizan, HR-Manager, Department of Laboratory services, Meenakshi
Mission hospital & Research center, Melur road, Madurai -625 107. And that no
part of the thesis has been submitted for the award of any other degree, diploma,
fellowship or other similar titles.

Place: Madurai G. Abisha

DATE: F. Angelin Jenit

R. Mari Selva Sundari

S. Maria Agnes Roganzia

T. Tvareta

N.S.Supraja

3
 ACKNOWLEDGEMENT

First and foremost we like to thank omnipotent almighty who showered his
choicest blessings and abounded grace, which helped in the successful
completion of our training in the Department of Laboratory Services,
Meenakshi Mission Hospital & Research Centre, Madurai.

We express our deep sense of gratitude to Dr. S. SaravanaSankar, Vice

Chancellor, KALASALINGAM UNIVERSITY, for providing us an
opportunity to enter into an esteemed institutes for training.

We express our grateful thanks to Dr. K. Sundar, Dean, School of Bio

and Chemical Engineering, and Dr. B. Vanavil, Associate Professor and Head of
the Department of Biotechnology, Kalasalingam University, Krishnankoil for
their support and valuable advice in carrying out of the research work.

We express our thanks to all the staff members in the Department of

Biotechnology, who were backbone for our success.

We would like to express our hearty thanks to Mr. Muthuchelizan, HR-

Manager, Department of Laboratory services, Meenakshi Mission Hospital &
Research Centre , Madurai who granted us permission to undergo a 15 days
training program in the reputed institution.

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 LIST OF CONTENTS

S.NO CONTENT PAGE

NO
1 MMHRC Profile
2 Introduction
3 Laboratory services
3.1 Blood collection
3.2 Hematology lab
3.3 Serology lab
3.4 Biochemistry lab
3.5 Histopathology lab
3.6 Microbiology lab
4 Result
5 Conclusion

5
 MMHRC PROFILE
Meenakshi Mission Hospital & Research Centre (SR Trust) in pursuit of medical
excellence has been delivering world class treatment care with an affordable cost with over 800
beds, MMHRC has growth to a multi-specialty hospital touching lives in and around Madurai.
We extend traditional Indian hospitality and International patients, combining it with our
cutting edge technology, clinical excellence and compassion to deliver quality health care to
all patients every single day. We also have shouldered responsibility and have pioneered
several charity initiatives such as informing people about the negative effects of smoking to
their body and encouraging them to try instead awaits you. We have fostered an environment
in which every person is motivated to continually improve the efficiency and effectiveness in
the management of health and services S.R. Trust is a non-profit organization. MMHRC prides
itself as its worthwhile contribution of having brought affordable and quality health care to the
people we embraced change and innovation, conforming to international standards, equipping
in the hospital with state-of-the-art technology, guided by experienced and dedicated doctors.

As we pass our 30th year we are proud to say that we have kept up the trust of
our patients in every possible way. With over 6 lakhs, patients treated so far, our relationship
with our patients has only strengthened. We began with the simple, yet profoundly demanding
mission of providing ‘world-class health care at affordable rates’. Our team of the doctors and
medical staffs have dedicated their learning, experience and time to this institution and to the
patients, who were willing to place their trust in our mission. Personal responsibility, team
work, and where necessary, collaboration with professionals beyond the hospital walls have
been the three key ingredients to our continual successes and growth. In keeping with this, we
have collaborated with Direct Relief International (DRI) to launch the Rare Diseases Project,
an endeavor that aims at treating patients of LSDSS (Lysosomal Storage Disorders Support
Society, India).

6
 INTRODUCTION

MMHRC which is registered under the Indian trust act (May 9, 1985). It was started in 1990
have made great achievements in the medical field by receiving awards and accolades ( Dr. B.
C. Roy, National Award ,Golden Peacock National Quality Awards ,Performance Excellence
Awards 2008).MMHRC Laboratory services has been functioning for over two decades,
forming the backbone of the hospital. The laboratory services offer more than 250 regular and
specialized tests. The test menu includes clinical and special chemistry, histopathology,
hematology, coagulation studies, serology, microbiology, endocrinology, tumor markers,
cardiac markers, anemic diseases, immunehistochemistry and chemilluminescence. Our
Laboratory services are backed by the best software available in the market for laboratory
services which is assisted by automated integration technology in order to maintain our
extracting standards of excellence and quality. Patients from various countries like Sri Lanka
and Maldives prefer our hospital for their medical treatment since we are able to offer the best
possible treatment for any disease comparable to any foreign country at a much cheaper cost.

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 DEPARTMENT OF LABORATORY SERVICES
A strong well established laboratory department quality assurance program with strict daily
two level internal quality control checks as well as monthly external quality control checks
with international and national comparison is the core of functioning of a department. The
laboratory department has the provision of standby equipment to prevent any delays in sample
processing in case of technical breakdowns. Instruments are bi-directionally interfaced to
improve work flow, reduced errors and prompt reporting to the clinical team. “Delivering
accurate results at lowest turnaround time with quality assurance checks is our goal”.
MMHRC which is an established hospital with huge volume of patients, will focus on top class
quality care at affordable cost.

Department comprises of six sections namely,

 Blood collection
 Hematology lab
 Bio-chemistry lab
 Serology lab
 Microbiology lab
 Histopathology lab

Blood collection:

MMHRC blood collection is a vast and sensitive department. It is responsible for

saving the lives of patients in critical situations, without any delay. Our essential functions are
donor motivation, blood collection from the appropriate donor, screening of donor blood,
processing and preparing the blood and blood components, proper storage of blood and blood
components and timely supply of blood components to the patients.

Hematology lab:

Hematology deals with diseases of blood, bone marrow and the lymph nodes. It includes
cancer like leukemia, a type of blood cancer; lymphoma, cancer affecting lymph glands and
myeloma, cancer of the immune cells. It is a branch in which molecular medicine is applied in
the day-to-day practice. Bone marrow transplantation life-saving treatment for many of the
hematological disorders and solid tumors.

Bio-chemistry lab:

Bio-chemistry deals with the routine diagnostic and specialized testing to some of the cardio-
vascular, diabetes, hormonal functions, transplant and oncologic physicians and surgeons. The
clinical bio-chemistry tests are performed with well-equipped art of instrumentation which is

8
capable of performing thousands of test per hour. New test and platforms are routinely
introduced to further both research and clinical trials.

Serology lab:

Serology lab analyzes blood specimens for diseases of public health significance. A
serology blood test is performed to detect and measure the levels of antibodies as a result of
exposure to a particular bacteria or virus. When people are exposed to bacteria or viruses
(antigens), their body’s immune system produces specific antibodies against the organism.
Anti-body levels (anti-body titer) help physicians determine whether an infection occurred
recently or years ago.

Microbiology lab:

It deals with microbes, this lab receives almost any clinical specimen, including swabs,
feces, urine, blood, sputum, cerebrospinal fluid, synovial fluid, as well as possible infected
tissue. The work here is mainly concerned with cultures, to look for suspected pathogens which,
if found, are further identified based on chemical tests. Also, sensitivity testing is carried out
to determine whether the pathogen is sensitive are resistant to a suggested medicine. Results
are reported which the identified organisms and the type and amount of drugs that should be
prescribed for the patients.

Histopathology lab:

Histopathology refers to the microscopic examination of tissues in order to study the

manifestation of diseases. Specifically, in clinical medicine, histopathology refers to the
examination of the biopsy or surgical specimen by a pathologist, after the specimen has been
processed and histological sections have been placed onto glass slides. In contrast, cyto-
pathology examines (1) free cells (2) tissue micro-fragments (as cell block).

BLOOD COLLECTION
The first step in acquiring a quality lab test result for any patient is the specimen collection
procedure. The venipuncture procedure is complex, requiring both knowledge and skill to
perform.

Procedure:

 Patient preparation
 Selecting the site
 Site preparation
 Tourniquet application and time
 Proper veni-puncture technique
 Order of draw
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  Proper tube mixing
 Correct specimen volume
 Proper tube handling and specimen processing
 Centrifugation
 Special handling of blood specimens
 Stability for whole blood, serum and plasma

Order of draw:

Blood collection tubes must be drawn in a specific order to avoid cross-contamination of

additives between tubes. The recommended order of draw for plastic vacutainer tubes is:

1. First-blood culture bottle or tubes

2. Second-coagulation tube
3. Third-non-additive tube
4. Last draw-additive tubes
 SST (red-grey or gold top)-contains a gel separator and clot activator.
 Sodium heparin (dark green top)
 PST (light green top)-contains lithium heparin anti-coagulant and a gel
separator.
 EDTA (lavender top)
 Oxalate/fluoride (light grey top) or other additives.

Note: Tubes with additives must be thoroughly mixed. Clotting or erroneous test results may
be obtained when the blood is not thoroughly mixed with the additive.

Labeling the sample:

All specimens must be received by the laboratory with a legible label containing at least two
(2) unique identifiers. The specimen must be labelled with the patient’s full name (preferably
last name first, then first name last) and one of the following:

 GHS medical record number (MRN) –for Geisinger locations, this is the
required second identifier.

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  Patient’s full date of birth (must include the month, day and year)
 Unique requisition identifier/label

Sample spillage cleaning procedure:

Display the sign board

Take the personal protective equipment

Mask, gloves, apron

Take a wet cloth and the clean the spilled sample

With SN7 gain clean the spilled area

Again with wet cloth clean the area

Then clean the area with wet cloth

Dispose the waste as per the bio-waste product

Do hand washing after the cleaning is over

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 HEMATOLOGY
Hematology is the study of the cells and proteins found in blood. A diagnostic hematology
laboratory is usually divided up into 4 main areas:

 Routine hematology – full blood examinations, morphology + other test.

 Coagulation – testing for the proteins and cells involved in clotting.
 Blood bank – blood and blood product transfusions.
 Special tests – performed only when required.

There are three main types of cells in our blood:

 Red blood cells (erythrocytes) transport oxygen around our body and
remove waste products and carbon dioxide.
 White blood cells (leucocytes) are responsible for fighting infections and
invading pathogen (e.g. viruses are bacteria).
 Platelets (thrombocytes) that help our blood clot if you are bleeding.

BLOOD COMPONENTS:

Blood differs from all of the other tissues in the body in that it has a large liquid component,
which gives it characteristic fluid properties. Like all other tissues, blood is composed of a
number of different cells, each with a unique function. These cells are erythrocytes (red blood
cells), leucocytes (white blood cells) and platelets. Because blood is the fluid, it has to be
contained in a closed system otherwise it would leak out in our body. This closed system is
called the vascular system (blood vessels).

Function of blood:

 Transport of oxygen.
 Transport of essential nutrients required for metabolism.

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  Transport of waste materials.
 Transport of cells involved in the defending the body against pathogens and toxins.
 Heat exchange mechanism.
These functions are all part of the process of homeostasis i.e., the maintenance of a
balanced steady state. Our heart is an extremely efficient pump, which is capable of re-
circulating our total blood volume every 10-12 minutes. In an average adult the blood volume
is approximately 5litres. Occasionally, the vascular system (blood vessels) may “spring a leak”
and blood will be lost, this is called bleeding or hemorrhage.
Blood is a tissue consisting of cells suspended in a fluid called plasma. The sample
of blood that has been collected into an anti- coagulant (substance to stop blood from clotting)
and then centrifuged to separate the red cells and plasma. The proportion of cells and plasma
in a healthy adult is indicated below although the amounts will vary slightly depending on the
age and sex of the individual. Adult males usually have more red cells than adult female.

Erythrocytes (red blood cells/RBC):

These appear at the bottom of the tube as they have the greatest density i.e., the
heaviest. Red blood cells are mainly involved in the transport of oxygen and removal of carbon
dioxide.

Buffy coat (leucocytes/WBC and platelets/PLT):

On top of the red cells is a greyish-white layer which is called a buffy coat. This layer
consists of the leucocytes or white cells and platelets. Leucocytes are part of the body’s defense
mechanism, while platelets are the cellular part of the hemostatic (clotting) system.

Plasma:

Above the buffy coat is a pale straw-colored fluid called plasma. Plasma contains
hundreds of different substance-proteins, vitamins, hormones, minerals. There are some
substances present in plasma, which are of interest to the hematology laboratory. These are the
coagulation proteins, which are involved in the clotting of blood.

Serum:

If blood is allowed to clot and then left for a while the clot will shrink and a fluid appears
above the clot. This fluid is called as serum. For some laboratory tests, serum is the specimen
of choice. Serum is almost the same as plasma except there are no clotting proteins present.

Collection of blood samples for testing in the laboratory:

Blood samples for testing are collected from the venous circulation using a needle
and syringe or the vacutainer system. The process of collecting blood from a patient is called
veni section. If a sample of blood is removed from the body and placed in a tube the blood will
eventually solidify; this process is called as clotting. Anti-coagulants can be used stop the
clotting process. The common anti-coagulants used are ethylene-di-amine-tetra-acetic acid
(EDTA), sodium citrate and heparin.

13
Ethylene-di-amine tetra-acetic acid (pink/purple top tube):

Ethylene-di-amine-tetra-acetic acid (EDTA) is a powder and blood collected into it does not
clot. On collection blood must be gently mixed so that the EDTA is dissolved in the blood.
EDTA prevents blood from clotting by binding or chelating (binding) the calcium ions from
the blood (calcium ions are necessary for blood to clot).Blood collected into EDTA is used for
routine hemotology tests.

Tri sodium citrate (light blue top tube):

Tri sodium citrate is a liquid anticoagulant that is used to collect blood for coagulation studies.
Samples must be gently mixed immediately after collection. Tri sodium citrate chelates calcium
ions from the blood (calcium ions are necessary for clotting).

Heparin (green top tube):

Heparin is an anticoagulant used for biochemistry tests and some specialized hematology test
Heparin prevents blood coagulation by inhibiting the action of thrombin. Thrombin is an
activated coagulation protein that converts fibrinogen to fibrin. Fibrin formation occurs when
the blood clots.

BIOCHEMISTRY LABORATORY

Biochemistry is a study of chemical process within and relating to living organisms. Much of
biochemistry deals with the structures, functions and interactions of biological
macromolecules, such as proteins, nucleic acids, carbohydrates and lipids, which provides the
structure of cells and perform many of the functions associated with life.

Recent past has seen major advances in diagnostic methodologies and

instrumentation(automation).

Purpose:

This is for technical persons involved in clinical biochemistry department to know about pre-
analytical analysis of sample (whole-blood, urine, serum, plasma) when received. Specimens
or samples are ANALYZED while substances in them are MEASURED AND
QUANTIFIED .Substances measured in serum fail generally into the following
categories:

Substances NORMALLY PRESENT with a function in the circulation

 Glucose
 TP
 Albumin
 Individuals proteins

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  Electrolytes
 TAG, Cholesterol
 Hormones
 Vitamins
Metabolites-nonfunctioning WASTE PRODUCTS in the process of being cleared
 Urea
 Creatinine
 Uric acid
 Ammonia
 Bilirubin
 Substances RELEASED FROM THE CELLS AS A RESULT OF CELL
DAMAGE AND ABNORMAL PERMEABILITY OR ABNORMAL
CELLULAR PROLIFERATION
 Enzymes such as LD,ALT,AST,CK,AMS,GCT,ALP,ACP
 FERRITIN
 Drugs and toxic substances:
 Antibodies
 Substances of abuse
 Therapeutic drugs
 Poisons

CLINICAL BIOCHEMISTRY TESTS:

The biochemical investigations (on blood/plasma/serum) carried out in clinical biochemistry

laboratory may be grouped into different types:

 BIOCHEMICAL PROFILES:

These tests are based on the fact that more useful information on the patient disease status
that can be obtained by analyzing more constituents rather than the one (e.g. plasma
electrolytes-Na+, K+, Cl-, bicarbonate, urea, liver function tests, serum, bilirubin, ALT,AST)

 DYNAMIC FUNCTION TESTS:

These tests are designed to measure the body response to external stimulus e.g;oral glucose
tolerance tests(to assess glucose hemostasis)bromo-sulphthfein to assess liver function tests).

 SCREENING TESTS:

These tests are commonly employed to identify the inborn errors of metabolism, and to check
the entry of toxic agents (pesticides, lead and mercury into the body).

 METABOLIC WORK-UP TESTS:

This is to identify the endocrinological disorders.

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 SPECIFICITY OF TESTS:

 RENAL FUNCTION-blood urea nitrogen (BUN), creatinine.

 ELECTROLYTE–Chloride, potassium, phosphorous, calcium, magnesium.
 LIVER- Alanine amino- transferase (ALT), aspiratate amino transferase (AST),
alkaline phosphatase, gamma-gluta-amyl transferase (GGT), total and direct bilirubin,
albumin.
 PANCREAS-Amylase, lipase, glucose.
 SERUM PROTEINS-Total protein, albumin, globulin, alpha, beta and gamma globulin
via protein electrophoresis.
 INJURY-Creatine kinase (CK), lactate de-hydrogenase (LDH).
 LIPIDS-Cholesterol, triglycerides, high-density lipoprotein (HDL), low- density
lipoprotein (LDL), very low- density lipo-protein (VLDL).

USES OF BIOCHEMICAL TESTS:

 DIAGNOSIS
 SCREENING THE DISEASE
 MONITORING PATIENTS
 RESEARCH STUDIES AND CLINICAL TRAILS OF NEW
DRUGS.

There are over 400 different tests which may be carried out in a clinical biochemistry
laboratories. They may vary from the very simple, such as measurements of sodium, to the
highly complex, such as DNA analysis, screening for drugs, or differentiation of lipoproteins
variants. Many high volume tests are done on large automated machines. Less frequently
performed tests may be conveniently carried out by using commercially prepared
reagentspackaged in “KIT” form. Some analyses are carried out manually.

SEROLOGY
Serology lab analyzes blood specimens for diseases of public health significance. A serology
blood test is performed to detect and measure the level of antibodies as a result of exposure to
a particular bacteria or virus. When people are exposed to bacteria or viruses (antigen), their

16
body’s immune system produces specific antibodies against the organism. Antibody levels
(antibody titer) help physicians determine whether an infection occurred recently or years ago.
Most of the serology tests are done by HEPA CARD.

Submitting specimens:

 Whole blood:
Refrigerated specimens must be received within 24 hours of the time of
collection.
 Serum separated from clot:
Refrigerated specimens must be received within 48 hours of the time of
collection. Frozen specimens must be received within six months of
collection.
 CSF:
Refrigerated specimens must be received either within 48 hours from the
time

1. PRIMARY SEROLOGICAL TESTS: (MARKER TECHNIQUES):

(e.g):

 Enzyme linked immuno sorbert assay (ELISA)

 Immuno fluroscent antibody technique(IFAT)
 Radio immuno assay(RIA)

2. SECONDARY SEROLOGICAL TESTS:

 Agglutination tests
 Complement fixation tests(CFT)
 Precipitation tests
 Serum neutralization tests(SNT)
 Toxin anti-toxin tests

3. TERTIARY SEROLOGICAL TEST :E.G.

 Determination of the protective value of an anti serum in an animal.

Antibodies are so diverse , various tests are useful for detecting the presence of different types:

 An agglutination assay shows whether antibodies exposed to certain antigens

will cause particles clumping.
 A precipitation test shows whether the antigens are similar by measuring for
the presence of antibody in the body fluid.
 The western blot test identifies the presence of antimicrobial antibodies in our
blood by their reaction with target reactions.

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 HEPACARD
HEPACARD is a visual, rapid, sensitive, and accurate one step immune assay for a
qualitative detection of hepatitis B surface antigen (HBs Ag) in human serum or plasma. The
assay is used as an aid in the recognization and diagnosis of acute infections and chronic
infections carrier of Hepatitis-B virus. It determines the specific HBV serotypes. Four antigenic
combinations such as adw, adr, ayw, ayr.

PRINCIPLE:

It is based on the sandwich principle. The monoclonal antibodies get conjugated

to colloidal gold and polyclonal antibodies. The thin line in the card contains an nitrocellulose
strip. The immunoassay is based on the antigen culture.

KIT CONTENTS:

 HEPA card test device

 Sample dropper
 Instruction manual

STORAGE:

The HEPACARD should stored in a temperature 2-30 C. It should be stored in

the coolest and driest area. It should not be frozen and must be protected from humidity.

LIMITATIONS:

 It should be used for in-vitro diagnosis use only.

 It should be treated with 0.5% of sodium hypochlorite for one hour.
 The card should be used only for 20 minutes only.
 Denaturation should be avoided.
 It should use for the detection of HBs Ag in serum and plasma only.

18
  Final positive results can be obtained due the presence of antigen or
elevated method levels of retention factor.
 If the HEPACARD is non-reactive with the clinical symptoms, additional
methods can be used.
 It is used for detecting diseases like HCV, TRI-DOT, NS3, NS4, NS5.
TEST PROCEDURE:

 Add 3 drops of buffer solution to the centre of the device.

 Hold the dropper vertically downwards and add one drop of patient sample (50*10-6
L of serum or plasma).
 Add 5 drops of buffer solution.
 Add 2 drops of protein-A conjugate.
 Read the result and discard the card immediately considering it to be potentially
infectious. .

MICROBIOLOGY LABORATORY
Microbiology is the study of microorganisms those being unicellular (single cell), multicellular
(cell colony) or acellular (lacking cells). Microbiology encompasses numerous sub-disciplines
including mycobacteriology, mycology and bacteriology. It includes bacteria, viruses, fungi,
prions, protozoa and algae and other microbes. It includes mycology, myco-bacteriology,
bacteriology.

INSTRUMENTS:

 Laminar flow chamber

 Automated analyser
 Loop sterilizer

Some of the commonly used agar are:

 Bile esculin agar:

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 It is used for differential isolation and presumptive identification of group D
streptococci in food and pharmaceuticals products.

 SS agar:
A differential selective medium for the isolation of salmonella and some
shigella species from pathological specimens suspended food stuffs etc.
 Potato dextrose agar:
It is used for the isolation and enumeration of yeasts and moulds from dairy
and other food products in accordance with FDA BAM 1998.
 Mannitol salt agar base:
It is used for selective isolation of pathogenic staphylococci.
 Brain heart infusion agar:
It is used for the cultivation of fastidious pathogenic bacteria, yeasts and
moulds.
 Hectoi enteric agar:
It is used for the isolation of salmonella and shigella from faecal specimen.
It contains lactose, sucrose and salicins.
 XLD agar:
Xylose lysine deoxycholate agar is a selective cum differential media which
is used for the isolation of enteric pathogen. The selective nature of the agar is increased
through the addition of deoxycholate.
 Chocolate agar:
It is used for the cultivation of fastidious microorganisms like H.influenzae,
N.meningitis, N.gonnorhea. This blood agar is heated at 100 degree celcius for 10
minutes. Additional enrichment and inhibitory substances are added to enhance the
growth of specific microorganisms.
 CLED agar:
Cystine lactose electrolyte deficiency medium with bromo thymol blue is
recommended for isolation, enumeration and identification of microorganisms.
 TCBS agar:

20
 In TCBS agar thymol blue and bromo thymol blue are included as indicators
of pH changes. It is used for the isolation and cultivation of VIBRIO. The alkaline pH
of the medium enhances the recovery of Vibrio cholera.

 Baired parker agar:

The selective agents are lithium chloride and potassium telluride. Egg yolk
emulsion is used to identify the proteolytic character of the isolate. It is used for the
isolation of coagulase positive Staphylococcus aureas.
 Cetrimide agar:
It is used for the isolation of Pseudomonas aeruginosa from pus, sputum,
drains etc., and also determining the ability of an organism to produce fluorescein and
pycocyanin.

HISTOPATHOLOGY

Histopathology is the study of tissues. It refers to microscopic examination of biopsy

or surgical specimen by a pathologist, after the specimen has been processed and
histological sections have been placed onto two glass slides. In contrast, cytopathology
(study of cells) examines free cells or tissue micro fragments (as “cell blocks”).

COLLECTION OF TISSUES:
Histopathological examinations of tissues starts with the surgery, biopsy or
autopsy. The tissue is removed from the body or plant and then often following expert
dissection in the fresh state placed in a fixative which stabilizes the tissue to prevent
decay. The most common fixative is formalin (10% neutral buffered formaldehyde in
water.

PREPARATION FOR HISTOPATHOLOGY:

 Sample receiving
 Fixation
 Crossing
 Tissue processing
 Embedding
 Section cuttings
 Staining
 IHC( Immuno Histochemistry Marker)

STAINING OF PROCESSED HISTOLOGY SLIDES:

21
 This can be done to slides processed by the chemical fixation or frozen section slides.
To see the tissue under a microscope, the sections are stained with one or more pigments. The
aim of staining is to reveal cellular components; counter stains are used to provide contrast.

Cross section of the tissue Staining of slides

The mostly commonly used stain in histopathology is a combination of hematoxylin

and eosin. Hematoxylin is used to stain nuclei blue, while eosin stains cytoplasm and the
extracellular connective tissue matrix pink. There are hundreds of various other techniques
which have been used to selectively stain cells. Other compounds used to colour tissue
selections include safranin, oil, red O, congo red and artificial cells. Histochemistry refers to
the science of using chemical reactions between laboratory chemicals and components within
tissue. A commonly performed histochemical techniques is the PERLS PRUSSIAN BLUE
reaction, used to demonstrate iron deposits in diseases like HEMOCHROMATOSIS.

Recently, antibodies have been to stain particular proteins, lipids and carbohydrates.
IMMUNOHISTOCHEMISTRY technique has greatly increased the ability to specifically
identify categories of cells under a microscope. Other advanced techniques include IN SITU
HYBRIDIZATION to identify specific DNA or RNA molecules. These antibody staining
methods often require the use of frozen section histopathology. These procedures above are
also carried out in the laboratory under scrutiny and precision by a trained specialist medical
laboratory science.

CONCLUSION

Technological in evolution clinical laboratory diagnostics has advanced considerably by

allowing for direct molecular detection of a pathogen in a clinical specimen rather than relying
on isolation and cultivation. This approach has decreased the time required to identify a
pathogen because the laboratory is no longer limited by the growth of kinetics of the organism.
Therefore, patients can be evaluated and if infected can be treated promptly, thereby
diminishing progression to disease and the disrupting transmission. As with the all the changes

22
in laboratory technology, a synthesis of scientific evidence is required the implementation of a
new or improved test platform.

23