Hematology Review

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 130,000-400,000. Active role in coagulation and hemostasis.

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

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

29. WBCEvaluation

30. MyelocyticMaturation 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 versusLeukomoid 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 MyelogenousLeukemia

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 Traumatic Tap 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 >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!!!!