1. Hematology Review By Felicia Magee Tardy, M.S., MT (ASCP)
Department of Clinical Pathology
University of Mississippi Medical Center
Jackson, Mississippi

2. Introduction Hematology: the study of blood and blood forming tissues.
Blood consists of 55% plasma and 45% formed elements.
Formed elements include erythrocytes, leukocytes, and thrombocytes.

3. Erythrocytes Normal range 4.2-5.5 million per mm3 in adults.
Biconcave shape.
Diameter 7 microns.
Cells for transport of O2 and CO2.
Life span 120 days.

4. Leukocytes Normal range 4 - 11 thousand per mm3 in adults. Five types.
Size 8-20 microns.
Involved in fighting infection, combatting allergic reactions, and immune responses.

5. Thrombocytes Smallest cells in the blood.
Normal range 130, ,000.
Active role in coagulation and hemostasis.

6. Anticoagulant of choice: EDTA
Routine Hematology
Anticoagulant of choice: EDTA
Complete Blood Counts (CBCs)
Manual WBC Differentials
Erythrocyte Sedimentation Rates (ESRs)
Sickle Screens
Reticulocyte Counts

7. Automated Counting Coulter Principle
Electrical impedance: resistance or change in current when cell passes between two electrodes in NaCl solution.

8. Automated Counting Flow Cytometry
Uses lasers to measure both forward and side scatter.
Forward scatter measures size.
Side scatter measures granularity.

9. Sources of Error Inadequate mixing of specimen. Hemolyzed specimens.
Lipemic specimens.
Cold agglutinins.
Clotted specimens.
Platelet clumps or platelet satellitosis.
Diluted specimens.

10. Know Normal Ranges!!! WBC RBC HGB HCT MCV MCH MCHC PLT MPV SEGS LYMPHS
MONOCYTES
EOSINOPHILS
BASOPHILS

11. Erythrocytic Maturation Series
Rubriblast
Prorubricyte
Rubricyte
Metarubricyte
Reticulocyte
Erythrocyte

12. RBC Morphology

13. Elliptocytes Target Cells

14. Tear Drops Stomatocytes

15. Sickle Cells Schistocytes

16. RBC Inclusions

17. Howell-Jolly Bodies Round, purple inclusions in RBCs. Composed of DNA.
Commonly seen in in patients with hypofunctioning spleens.
Splenectomy.

18. Basophilic Stippling Numerous, small purple inclusions in RBCs.
Aggregates of ribosomal RNA.
Most commonly seen in lead poisoning.

19. Pappenheimer Bodies
Clusters of dark blue granules, irregular in size and shape.
Composed of iron and ribosomal RNA.
Seen in sideroblastic and hemolytic anemias.

20. Classifications of Anemias
Microcytic, Hypochromic
Iron deficiency
Sideroblastic
Chronic disease, Inflammation
Lead poisoning
Thalassemia trait

21. Microcytic, Hypochromic
Many RBCs smaller than
nucleus of normal
lymphocytes, increased central pallor.
Iron deficiency, thalassemias, anemia of chronic disease.

22. Classifications of Anemias
Normochromic
Hereditary Spherocytosis
Hereditary Elliptocytosis
PNH
G6PD deficiency
Aplastic anemia
Acute blood loss

23. Classifications of Anemias
Macrocytic
Vitamin B12 deficiency
Folate deficiency
Liver disease

24. Macrocytic RBCs Most RBCs larger than nucleus of normal
lymphocytes, increased MCV.
Folate or Vitamin B12 deficiencies, alcoholism, and liver disease.

25. Reticulocytes Immature RBCs. Contain residual ribosomal RNA.
Reticulum stains blue using a supravital stain (new methylene blue).
Counted and expressed as % of total red cells.

26. Reticulocyte Count
Uses supravital stain which stains cells in the living state.
Retic % = # retics per 1000 RBCs 10
Corrected retic= % retics x pt. HCT 45

27. Beta Chain Substitutions
Hemoglobinopathies
Beta Chain Substitutions
Hgb S: Valine for glutamic acid
(6th position, beta chain)
Hgb C: Lysine for glutamic acid

28. Alkaline Electrophoresis
Hemoglobinopathies
Alkaline Electrophoresis
- C S F A +
A2 D
E G O

29. WBC Evaluation

30. Myelocytic Maturation Series
Myeloblast
Promyelocyte
Myelocyte
Metamyelocyte
Band Neutrophil
Segmented Neutrophil

31. Toxic Granulation Increased basophilic granules in neutrophils.
Seen in severe infections, burns, malignancies, and pregnancy.
Distinguish from basophils.

32. Dohle Bodies Sky blue inclusions in cytoplasm of neutrophils.
Seen in infections, burns, myleproliferative disorders, and pregnancy.
Composed of RER and glycogen granules.

33. Pelger-Huet Anomaly

34. Special Stains PAS: ALL, Erytrholeukemia
LAP: CML v. Leukomoid Reaction
Myeloperoxidase: AML, AMMoL (weak+)
Sudan Black: AML, AMMoL (weak+)
Specific Esterase: AML, AMMoL
Non-specific Esterase: AMML, AMoL
TRAP: Hairy Cell Leukemia
TDT: ALL
Prussian Blue: Sideroblastic Anemia

35. CML versus Leukomoid Reaction
Characteristic CML Leukomoid
LAP score Decreased Increased
Toxic gran Decreased Increased
Dohle bodies Absent Present
Philadelphia May be present Absent
chromosome

36. WBC Disorders
Alder-Reilly: Large azurophilic granules, increased mucopolysaccharides
Chediak-Higashi: Membrane defect of lysosomes, large primary granules in segs
May-Hegglin: Familial disorder with large platelets and Dohle bodies
Pelger-Huet: Hyposegmented neutrophils

37. FAB Classifications M1: Myeloblastic without maturation
M2: Myeloblastic with maturation
M3: Promyelocytic
M4: Myelomonocytic
M5a: Monocytic, poorly differentiated
M5b: Monocytic, well differentiated
M6: Erythroleukemia, DiGuglielmo’s
M7: Megakaryoblastic

38. Acute Myeblastic Leukemia (M1)

39. Chronic Myelogenous Leukemia

40. FAB Classifications L1: Small, uniform lymphoblasts
L2: Large, pleomorhphic lymphoblasts
L3: Burkitt’s type (vacuolated and
deeply basophilic cytoplasm)

41. Acute Lymphoblastic Leukemia

42. Chronic Lymphocytic Leukemia

43. Body Fluids

44. Types of Body Fluids Cerebrospinal Fluid (CSF) Pleural Fluid
Pericardial Fluid
Peritoneal Fluid
Synovial Fluid
Amniotic Fluid
Miscellaneous Fluids

45. Body Fluids Testing Physical examination RBC and WBC counts
WBC differential
Crystal examination*
Rope’s test*
*Refers to synovial fluids only.

46. Cerebrospinal Fluid (CSF)
Examined to determine the presence of meningitis or intracranial hemorrhage.
Multiple tubes drawn (4).
Differentiate between traumatic tap and intracranial hemorrhage.
Xanthochromia is indicative of intracranial hemorrhage.

47. Quick Review
Differentiate between traumatic tap and intracranial hemorrhage.
Differentiate between transudate and exudate.
Crystal identification and associated conditions.

48. Traumatic Tap v. Intracranial Hemorrhage
Decreasing amounts of blood with each tube drawn
May contain clots
No xanthochromia
Supernatant clear
Intracranial Hemorrhage
Blood evenly distributed throughout collection tubes
No clots
Xanthochromia
Hemosiderin, hematoidin crystals

49. Transudates v. Exudates
Characteristic Transudate Exudate
Appearance Clear, colorless Yellow, turbid, purulent, bloody
Specific gravity < >1.015
Protein <3 g/dL >3 g/dL
LD <200 IU >200 IU
Cell count <1000/uL >1000/uL
Conditions Congestive Infections,
Heart failure Malignancies

50. Crystal Identification
Monosodium urate (uric acid)
Yellow when parallel to the compensator, and blue when perpendicular.
Seen in gout.
Calcium pyrophosphate
Blue when parallel to the compensator, and yellow when perpendicular.
Seen in pseudogout.

51. Crystal Identification

52. Good Luck!!!!