ADVANCED HEMATOLOGY LAB

OBJECTIVES/Outline
At the end of this introduction to clinical laboratory hematology
course/ semester, the student will be able to state these
principles of clinical hematology lab procedures and perform the
procedures. All differential values, WBC estimates, and platelet
estimates, reticulocyte counts, manual hematocrit, and other lab
values reported by the student must match the automated CBC/
or technologist/lab reported values within 20%.

Psychomotor domain objectives are denoted in red ink. Cognitive

domain objectives are denoted in black ink.
Section 1.
1. Perform a peripheral blood smear review
State the procedures and observations performed under
each objective (magnification) and perform the
procedures
a. Observations under 10X
i. Observe the slide. Check for slide quality, platelet
clumping, WBD distribution, and unusual cells.
ii. Locate a good area to begin counting.
b. Observations under 40 X
i. Perform a WBC estimate, showing the calculations.
ii. Report the WBC estimate to the correct number of
significant figures and in the correct units.
iii. Compare WBC estimate to WBC count reported by
automated analyzer; estimated results must
match reported results within 20%
c. Observations under 100 X
i. Perform a 100 WBC differential and report relative
counts.

2.
3.
4.
5.
6.

7.

ii. If > 5NRBCs are present in a 100 WBC count

differential, correct the WBC count for the
presence of NRBCs.
iii. Read and grade RBC morphology, by the RBCM
grading criteria; include significant RBC
morphology and RBC inclusion bodies.
1. Compare your results to the results that the
laboratory technologist reported; results
should match within one grading scale.
iv. Perform a platelet estimate; show your calculation.
v. Report the platelet estimate with the correct
number of significant figures and in the correct
units.
vi. Compare your platelet estimate with the platelet
count reported by the automated analyzer; results
should match within 20%.
vii. Report a platelet adequacy comment, according to
grading criteria based on comparison with platelet
count reference ranges.
d. State which objectives use oil immersion and which do
not.
State the procedure and perform a platelet estimate.
State the platelet estimate calculation.
Perform a platelet estimate on a peripheral blood smear, under
100x oil immersion objective .
State the procedure and perform a WBC estimate. State and
use the WBC estimate calculation.
Describe the pattern of reading a peripheral smear when
counting and differentiating cells. What is the pattern called
a. Demonstrate the battlement pattern of reading a PBS
and use it to read differentials and reticulocyte counts.
Identify the correct area of a blood smear in which to evaluate
RBC Morphology, to count platelet estimates and WBC
estimates, and perform a complete WBC differential.

Section 2.

1. Describe and perform the procedure for performing a manual

reticulocyte count.
P-2
a. Make a reticulocyte stain preparation using EDTA whole
blood and New Methylene blue supravital stain, with the
correct dilution and incubation time.
b. Make a high quality PBS of the stained blood cells.
c. Read the reticulocyte count, without a Miller ocular, by
counting the number of reticulocytes seen in 1000
RBCs.
d. Calculate the % reticulocytes; show calculations.
e. Report the reticulocyte count, in the appropriate units;
results must match reported value to within 20%.
2. Define and recognize the morphology of a reticulocyte when
stained with a supravital stain. What content of the RBCs are
precipitated with supravital stains.
3. Name two supravital stains used in a manual reticulocyte
count.
4. State whether reticulocytes be seen when the slide is stained
with Wrights stain. If so, describe their appearance.
5. Describe how to perform a reticulocyte count without a Miller
Ocular.
a. Calculate the % reticulocytes using the calculation for
retic counts by the Miller ocular method; show your
calculations.
b. Report your reticulocte count and compare to the
reported retic count; results must match within 20%.
6. Write and use the formulas to calculate
a. Relative reticulocyte count
b. Absolute reticulocyte count
c. Corrected reticulocyte count for anemia (low HCT)
d. Reticulocyte Production Index (RPI)
7. State the reference values for adults for reticulocyte counts
and the units.
8. Describe how the reticulocyte count and the RPI are used to
monitor anemia and its therapy effectiveness.

9. Use the fine focus to differentiate reticulocytes from stained

artifacts to minimize error in estimating reticulocyte counts.
P-3
1. State the rule of three (RO3) related to RBC, HGB, and HCT
relative concentrations. Evaluate results to see if they
comply with the RO3.
2. Define and describe the 3 RBC Indices: MCV, MCH, MCHC
3. Provide the formula for MCV, the units, and the reference
values
4. Provide the formula for MCH, the units, and the reference
values
5. Provide the formula for MCHC, the units, and the reference
values
6. Calculate the MCV, MCH, and MCHC given the RBC count,
HGB, & HCT P-2
Section 6.
1. State and use the calculation of a Corrected WBC Count for
the presence of Nucleated RBCs (NRBCs)
2. State how many NRBCs need to be present (criteria) to
correct the WBC for the presence of NRBCs.
3. State whether a platelet estimate can be performed and
reported if platelet clumps are present on the slide? Why or
why not?
4. Manual estimates of platelets and WBCs should match the
automated counts by what % ?
5. Describe the procedure for performing a differential. Include
the sequence/order of examining the slide to review all the
necessary components of the differential. State all
components that should be reviewed for each cell
component.
6. Provide (and perform) the calculation to calculate an
absolute count from a relative count.
7. Recognize, identify, and label RBC morphology, and state the
conditions in which they are seen

8. Identify the following cells by their MCV

P-2
a. Normocytes
b. Microcytes
c. Macrocytes
9. Classify cells by their size and chromicity based on given
MCV and MCHC levels.
Section 8. More RBC Morphology
1. Recognize RBC Morphology, including RBC inclusion bodies,
and identify cells by name and diseases by patterns of
abnormal RBC morphology. P-3
2. Grade RBCM by grading protocol. Results must match the
results reported by the lab technologist by a maximum of
one grading scale difference P-2
Section 12.
Section 13. Laboratory safety guidelines
1. State the dress code in a laboratory
a. Dress according to dress code
2. Describe universal precautions.
a. Wear personal protective equipment at all times in
lab. P-1
AFFECTIVE OBJECTIVES:
At the end of this course, the student should be able to
1. Recognize unacceptable results and correct problems, when
possible.
2. Observe all established safety procedures and universal
precautions.
3. Maintain a clean and orderly work area.
COGNITIVE LEARNING OBJECTIVES FOR INTRODUCTION TO
THE CLINICAL LABORATORY
1 List examples of biological and chemical hazards and appropriate
laboratory safety guidelines.
2. Define: universal precautions, aseptic technique, biological safety cabinet

COGNITIVE LEARNING OBJECTIVES FOR PHLEBOTOMY:

At the end of this course, the student should be able to
1. Explain the purpose and practice of universal precautions.
2. Name the anticoagulant associated with the following color- coded tubes:
lavender.
3. Explain how the following anticoagulants prevent clotting: EDTA.
4. Give examples of preanalytical, analytical and postanalytical errors.
5. Discuss the proper disposal of used equipment and supplies.
COGNITIVE LEARNING OBJECTIVES FOR INTRODUCTION TO
HEMATOLOGY:
At the end of this course, the student should be able to
1. List the various types of anticoagulants and the appropriate use of each.
2. Describe the morphology and function of normal mature erythrocytes.
3. Define the following parameters and explain how they are derived:
a) rbc count; b) hematocrit ; c) MCV; d) MCH; e) MCHC.
4. List the reference ranges for RBCs, hematocrit, MCV, hemoglobin, MCH, and
MCHC in
a) adult males; b) adult females; c) infants; d) newborns
5.. Describe the morphology and function of normal mature leucocytes.
6. Differentiate WBCs: neutrophils, monocytes, lymphocytes, eosinophils and
basophils, bands, metamyelocyte, myelocyte.
7. State the reference ranges for the various leucocytes and explain the significance
of neutrophilia and lymphocytosis.
8. Distinguish between absolute counts and relative counts. Calculate absolute
WBC counts for subsets of WBCS and reticulocytes when given total WBCs and
Total RBC count.
9. Describe the morphology and function of mature platelets.
10. Explain the consequences of increased and decreased numbers of platelets.
11. Discuss the significance of polychromatophilia and an increased reticulocyte
count.
12. Define, recognize, and grade the following RBC morphpologies : anisocytosis,
microcytosis, macrocytosis.
13. Explain the formula and application for the RDW value.
14. Define the following terms: poikilocytosis, sickle cells, target cells, burr cells,
hypochromia.
15. Summarize the changes that occur during erythrocyte maturation.
1. Characterize the morphology of the RBCs in the stained smear, indicating
size, shape and staining patterns.

2. Recognize immature erythrocytes and leukocytes on a stained smear.

3. Correctly estimate the number of platelets on a stained smear and
characterize the morphology.
4. Perform a manual reticulocyte count using a vital staining technique.
5. Given the hematocrit and RBC count, calculate correctly the MCHC
and the MCH.
6. Read a complete differential.
7. Stain the blood smear with Giemsa or Wright stain.
1. Discuss safety in the lab
2. Describe proper use and care of the microscope
3. Review of staining procedure and proper differential evaluation

1. Perform normal peripheral blood smear differentials

2. Perform reticulocyte count
1. Perform normal peripheral blood smear differentials
2. Evaluation of peripheral blood smears for anemia (IDA and MBA)
1. Perform WBC and platelet counts and estimates
2. Describe methods to detect and monitor anemia (HGB, RETIC, Absolute
RETIC, RPI, Microhematocrit
1. Describe lab tests that indicate RBC destruction
2. Calculate RBC indices
3. Calculate reticulocyte and RPI