ADVANCED HEMATOLOGY LAB OBJECTIVES/OutlineAt 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 reviewState the procedures and observations performed under each objective (magnification) and perform the procedures a. Observations under 10Xi. 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 Xi. 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 Xi. Perform a 100 WBC differential and report relative counts.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.2. State the procedure and perform a platelet estimate.3. State the platelet estimate calculation. 4. Perform a platelet estimate on a peripheral blood smear, under 100x oil immersion objective .5. State the procedure and perform a WBC estimate. State and use the WBC estimate calculation. 6. Describe the pattern of reading a peripheral smear when counting and differentiating cells. What is the pattern calleda. Demonstrate the battlement pattern of reading a PBS and use it to read differentials and reticulocyte counts.7. 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-2a. 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 calculatea. Relative reticulocyte countb. Absolute reticulocyte countc. 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, MCHC3. Provide the formula for MCV, the units, and the reference values4. Provide the formula for MCH, the units, and the reference values5. Provide the formula for MCHC, the units, and the reference values6. Calculate the MCV, MCH, and MCHC given the RBC count, HGB, & HCT P-2Section 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 seen8. Identify the following cells by their MCVP-2a. Normocytesb. Microcytesc. Macrocytes9. Classify cells by their size and chromicity based on given MCV and MCHC levels.Section 8. More RBC Morphology1. Recognize RBC Morphology, including RBC inclusion bodies, and identify cells by name and diseases by patterns of abnormal RBC morphology. P-32. Grade RBCM by grading protocol. Results must match the results reported by the lab technologist by a maximum of one grading scale difference P-2Section 12.Section 13. Laboratory safety guidelines1. State the dress code in a laboratorya. Dress according to dress code2. Describe universal precautions.a. Wear personal protective equipment at all times in lab. P-1AFFECTIVE OBJECTIVES:At the end of this course, the student should be able to1. 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 LABORATORY1 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 to1. 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 to1. 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) newborns5.. 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 lab2. Describe proper use and care of the microscope3. Review of staining procedure and proper differential evaluation

1. Perform normal peripheral blood smear differentials2. Perform reticulocyte count

1. Perform normal peripheral blood smear differentials2. Evaluation of peripheral blood smears for anemia (IDA and MBA)

1. Perform WBC and platelet counts and estimates2. Describe methods to detect and monitor anemia (HGB, RETIC, Absolute RETIC, RPI, Microhematocrit

1. Describe lab tests that indicate RBC destruction2. Calculate RBC indices3. Calculate reticulocyte and RPI