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Peripheral Blood

Blood-Hematopoiesis-Lymphatics. Peripheral Blood. William F. Kern, M.D. Director, Laboratory Hematology Department of Pathology william-kern@ouhsc.edu.

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Peripheral Blood

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  1. Blood-Hematopoiesis-Lymphatics Peripheral Blood William F. Kern, M.D. Director, Laboratory Hematology Department of Pathologywilliam-kern@ouhsc.edu

  2. Downloading or copying any of the photographs, images or diagrams from this presentation for any purpose other than studying for BHL is prohibited

  3. Blood: Constituents • Plasma (liquid) • Cells: • Erythrocytes (Red Blood Cells) • Leukocytes (White Blood Cells) • Platelets

  4. Anticoagulated Blood Sample Allowed to Settle Plasma (50-60%) T Buffy Coat (WBCs & Platelets) R Erythrocytes (40-50%) R T Packed Cell Volume (“Hematocrit”) = x 100 [%]

  5. Plasma • Plasma = Liquid phase of anticoagulated blood • ~90% Water • Proteins: • Albumin • Coagulation factors • Immunoglobulins, lipoproteins, others • Ions, glucose, amino acids, waste products Note: Serum = fluid remaining after blood clotted

  6. ErythrocytesRed Blood Cells (RBCs) • Primary function: Oxygen transport • Hemoglobin = Oxygen carrying molecule • Oxygen bound to iron in heme ring • Secondary function: Acid-base balance • Anucleate • Life span ≈ 120 days • Most phagocytized & destroyed in spleen

  7. Hemoglobin • Heme = iron atom in protoporphyrin ring • Globin = protein portion • Hemoglobin = tetramer: 2 a-chains, 2 b-type chains: • Hb A: a2b2>95%* • Hb A2: a2d2 1.7-3.3%* • Hb F: a2g2 <2%* * Normal adult values at OUMC

  8. Hemoglobin Molecule

  9. ErythrocytesRed Blood Cells (RBCs) • Reddish-orange color on blood smear • Biconcave disk shape • Central pallor: ~ 1/3rd diameter of RBC • Diameter: ~ 7-8 microns • ~ Same size as nucleus of small, resting lymphocyte

  10. Erythrocytes Erythrocyte Erythrocyte Lymphocyte

  11. Erythrocytes: Descriptive Terms • Normocytic: Normal size • Microcytic: Small (<6.5 microns) • Macrocytic: Large (>8.5 microns) • Normochromic: Normal central pallor • Hypochromic: Widened central pallor (>1/2 diameter of cell)

  12. Reticulocytes • Reticulocyte = immature RBC containing RNA • Contain RNA for ~1 day after leaving marrow • Normal reticulocyte count ~ 1% of RBCs (~1% of RBCs replaced daily) • Basophilic (bluish) on routine blood smear stains • Definitive identification requires special stain (“reticulocyte stain”)

  13. Reticulocytes Reticulocyte

  14. Erythrocytes: Normal Parameters (Note: Values vary between different laboratories) MCV = Mean Corpuscular Volume (Femtoliters [10-15 L])

  15. Leukocytes:White Blood Cells (WBCs) • Granulocytes: Contain specific granules • Neutrophils (polymorphonuclear leukocytes or “Segs”) • Eosinophils (red granules) • Basophils (dark blue granules) • Agranulocytes: May contain lysosomes (nonspecific granules): • Lymphocytes • Monocytes

  16. Neutrophils (“Polys”) • Most common type of leukocytes • Function: Phagocytosis, predominantly bacteria • Appearance: • Nucleus segmented (“Seg”) or horseshoe-shaped (“Band”) • Granules small, weakly stained (“neutrophilic”) Neutrophils are your main defense against bacterial infection

  17. Segmented Neutrophil(PMN; “Seg” or “Poly”) • Nucleus with distinct lobes connected by thin strands of chromatin • Finely granular cytoplasm • Weakly staining granules

  18. Band Neutrophil (“Band”) • Non-segmented, horseshoe-shaped nucleus • Less mature than segmented neutrophil • Fully functional • Count towards absolute neutrophil count

  19. Neutrophils (“Polys”) • Lifespan: ~10 hours in circulation, 1-4 days in tissue • Increase in acute infection, inflammation or other stress: • Increase in “band” forms particularly suggests acute infection or inflammation

  20. Eosinophils (“Eos”) • Large, intensely red (“eosinophilic”) granules • Function: Phagocytosis of antigen-antibody complexes; kill parasites • Normally ~ 2-4% of WBCs: • Eosinophil number increase in allergic reactions & parasitic infections • Lifespan in blood ~ 8 hours

  21. Eosinophils (“Eos”) • Segmented nucleus- usually bilobed • Large, uniform, reddish cytoplasmic granules

  22. Basophils (“Basos”) • Large, dark blue (“basophilic”) granules • Least common leukocytes (≤1%): • Increase in chronic infections; some neoplasms • Function not entirely clear; may control immune reactions • Possibly related to tissue mast cells

  23. Basophil (“Basos”) • Large, purple granules • Segmented nucleus

  24. Lymphocytes (“Lymphs”) • Nucleus round; ~size of normal RBC • Usually scanty, pale blue cytoplasm • Normally ~20-40% of WBC (higher in children): • Increase in viral infections, Toxoplasmosis, other conditions • Infectious mononucleosis: Reactive or “atypical” lymphocytes

  25. Lymphocytes (“Lymphs”) • Small, round, condensed nucleus • Scant pale blue cytoplasm • Nucleus ~ same size as erythrocyte Small resting lymphocyte

  26. Lymphocytes (“Lymphs”) • May recirculate between blood and tissue • Divided into two main types: • B-Cells: Humoral immune system • T-Cells: Cell-mediated immunity; Control of immune system Natural killer cells = 3rd type of lymphocyte

  27. Lymphocytes: B-Cells • B-Cell = Bursa of Fabricius in birds; Bone marrow in humans: • Early maturation & differentiation in bone marrow • Final maturation after stimulation by antigen in tissues • Differentiate into plasma cells: Antibody synthesizing cells

  28. Lymphocytes: T-Cells • T-Cell = Thymus • Originate in bone marrow • Migrate to thymus for differentiation and maturation • Key regulation & control of immune system • Effector cells of cell-mediated immunity

  29. Lymphocytes: T-Cells • Two Main Types: • T-Helper: Express CD4 antigen; Master control cells of immune system • T-Suppressor: Express CD8 antigen Note: The HIV virus preferentially infects CD4+ lymphocytes by recognition of CD4 molecule on surface of T-cell

  30. Large Granular Lymphocyte • More cytoplasm • Large “azurophilic” (reddish) granules • ~10-15% of blood lymphocytes • Associated with natural killer function

  31. Reactive (“Atypical”) Lymphocytes • Characteristic of infectious mononucleosis (acute EBV infection) • May occur in cytomegalovirus (CMV) and other infections • Features: • Abundant, pale-blue cytoplasm • Larger, less condensed nucleus; may have nucleolus • Characteristically “hug” or “skirt around” erythrocytes

  32. Reactive (“Atypical”) Lymphocytes

  33. Monocytes (“Monos”) • Folded or bean-shaped nucleus • Abundant light grey to pale blue cytoplasm • May have small cytoplasmic granules (lysosomes) • ~3-8% of WBCs

  34. Monocytes (“Monos”) • Folded nucleus • Abundant, light grey cytoplasm • Cytoplasmic vacuoles common • Few, small cytoplasmic granules

  35. Monocytes • Circulate in blood for ~8-14 hours • Long survival in tissues • May differentiate into tissue macrophages or histiocytes

  36. Monocytes (“Monos”) • Two main functions: • Phagocytosis: Organisms (fungi, mycobacteria), debris, foreign material • Antigen processing and presentation to lymphocytes: • Antigen presenting cells are specialized; no phagocytic function • Examples: Langerhans cells in skin

  37. Leukocyte DifferentialProportion of Different WBC Types (Adult) Mnemonic: Never Let Monkeys Eat Bananas

  38. Leukocyte DifferentialAbsolute Numbers (OUMC; Adult)* *Ranges vary between different labs and between children & adults

  39. Platelets (Thrombocytes) • Smallest cellular elements: ~1-4 m diameter • Anucleate; light blue with reddish-purple granules • Adhere to injured blood vessel wall: • Immediate hemostatic plug • Form surface for coagulation cascade

  40. Platelets (“Thrombocytes”)

  41. Platelets: Platelet Count • Normal number: ~140,000-440,000 per mL (140-440 x 109/Liter)* • >20,000/mL: Adequate hemostasis for normal conditions • >50,000-100,000/mL: Adequate for most surgery • <10,000/mL: Risk of spontaneous hemorrhage * Varies in different laboratories

  42. Review

  43. What are the arrowed objects? What is their function?

  44. What is this? • What is its function?

  45. What is the arrowed object?What is its function?

  46. What is the arrowed object?

  47. What is the arrowed object?What is its function?

  48. Complete Blood Count (CBC) • Red Cells: RBC count, hemoglobin, hematocrit: • Red cell indices: MCV, MCH, MCHC • Red cell distribution width (RDW) • White Cells: Total WBC count: • Differential: % and absolute number • Platelets:Platelet count • Mean platelet volume (MPV) Most important values in yellow/bold

  49. Hematology Analyzers: Units 1 x 106/mL = 1 x 1012/L 1 x 103/mL = 1 x 109/L 1 gram/dL = 10 grams/Liter

  50. Automated Hematology Analyzer:White Blood Cells • Hematology analyzer reports 5-part differential: • Neutrophils, lymphocytes, monocytes, eosinophils, basophils • Percent and absolute number usually given • “Band” neutrophils included in manual differential only • Abnormal or immature white cells “flagged” so smear is made & examined

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