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Edit Comment Close Premium member Presentation Transcript Slide 1: CYTOLOGICAL EXAMINATION OF BONEMARROW Peripheral blood cells : Peripheral blood cells Bonemarrow cells : Bonemarrow cells Leukemia cells : Leukemia cells Lymphocytic leukemia cells : Lymphocytic leukemia cells Source of blood cells? : Source of blood cells? ？ ？ Slide 12: ERYTHROCYTE SERIES Normoblast Basophilic normoblast : Normoblast Basophilic normoblast polychromatic normoblast ortho-chromatic normoblast : polychromatic normoblast ortho-chromatic normoblast Net erythrocyte Erythrocyte : Net erythrocyte Erythrocyte Slide 16: GRANULOCYTE SERIES Myeloblast Promyelocyte : Myeloblast Promyelocyte Metamyelocyte : Metamyelocyte Neutrophilic myelocyte Band granulocyte : Band granulocyte segmented granulocyte Eosinophilic myelocyte : Eosinophilic myelocyte Metamyelocyte Eosinophilic band granulocyte : Eosinophilic band granulocyte Eosinophilic segmented granulocyte Basophilic myelocyte : Basophilic myelocyte Metamyelocyte Basophilic segmented granulocyte : Basophilic segmented granulocyte Basophilic band granulocyte Slide 24: LYMPHOCYTE SERIES Lymphoblast : Lymphoblast Prolymphocyte Lymphocyte Slide 26: MONOCYTE SERIES Monoblast : Monoblast Promonocyte Monocyte Slide 28: PLASMA SERIES Plasmablast : Plasmablast proplasmacyte plasmacyte Slide 30: MEGAKARYOCYTE SERIES Megakaryoblast : Megakaryoblast promegakaryocyte Granular megakaryocyte : Granular megakaryocyte Thrombocytogenous megarkaryocyte Naked nucleus : Naked nucleus Platelet Other cells : Other cells Organization of Alkaliphilic cells : Organization of Alkaliphilic cells Fibroblasts Mesh phagocytes : Mesh phagocytes Fat cells Endothelial cells : Endothelial cells Osteoblasts Osteoclasts : Osteoclasts Bone Marrow -Definition : Bone Marrow -Definition •“Bone” refers to location ofbone marrow after birth •“Marrow”: Latin for “middle”. •Hence Bone Marrow is locatedin middle of bone. DISTRIBUTION OF BONE MARROW : DISTRIBUTION OF BONE MARROW •Pelvis-40% •Vertebrae-28% •Cranium-mandible-13% •Ribs-8% •Sternum-2% •Ends of long bones-8% TYPES OF MARROW : TYPES OF MARROW Yellow marrow (inactive) composed primarily of fat Red marrow (active in hematopoiesis) Slide 44: Peripheral blood CBC Differential count Reticulocyte count Blood smear morphology Bone marrow aspirate Morphology cytochemical stains Flow cytometric immunophenotyping cytogenetic analysis Molecular biological studies BONE MARROW EXAMINATION - Contents Slide 45: Bone marrow tissue section Trephine biopsy Clot section Laboratory studies-miscellaneous Iron studies B12/folate Hemoglobin electrophoresis Immunoelectrophoresis LDH Coomb's test Microbiology culture/serology results Miscellaneous Indications For Bone Marrow Evaluation : Indications For Bone Marrow Evaluation Evaluation for malignancy Primary hematologic disorder Staging for Hodgkin's disease Staging for non-Hodgkin's lymphoma Metastatic tumor (nonhematopoietic) Monitoring therapy Postchemotherapy Post bone marrow transplantation Evaluation of Erythropenias Marrow production problem Peripheral destruction inadequate/ ineffective marrow release Miscellaneous Culture (fever of unknown origin) BM SITES AND INDICATIONS FOR ASPIRATION : BM SITES AND INDICATIONS FOR ASPIRATION Posterior iliac crest (any age, for aspiration and biopsy) Anterior iliac crest (only for infants and children, for aspiration and/or biopsy) Sternum (unsuitable under 15 y/o;only for aspiration) Tibia (infants below 18 mo.; only foraspiration) Slide 48: X- sternal aspiration sites Tibial aspiration site Posterior iliac crest aspiration Marrow puncture needle - Salah : Marrow puncture needle - Salah Scheme for examining BM aspirate films : Scheme for examining BM aspirate films 1) Determination of cellularity 2) Identification of megakaryocytes- Note morphology and maturation sequence 3) Look for clumps of abnormal cells 4) Identify macrophages Examine for evidence of haemophagocytosis , malaria pigment, bacterial or fungal inf (x 40 , x 100) 5) Identify all stages of maturation of myeloid and erythroid cells 6) Determine myeloid:erythroid ratio 7) Perform a differential count 8) Look for areas of BM necrosis 9) Assess the Iron content of macrophages Slide 51: Normal cellularity Hypocellular Hypercellular Staining of cells : Staining of cells Identification of various types of blood cells Vacuoles : Vacuoles Toxic granules Lymphocytic leukemia : Lymphocytic leukemia Microfilaria : Microfilaria Plasmodium Leishmania - Amastigotes : Leishmania - Amastigotes Toxoplasma - Bradyzoites Staining methods : Staining methods POX、α-NAE、NAP、PAS、Fe dyes (Peroxidase, POX) : (Peroxidase, POX) Principle: Benzidine+ H2O2 [O] Aniline blue + Nitroso sodium cyanide Granules (azurophilic) are stained Cells of Myeloid series - POX Positive Cells of lymphocytic, megakaryocytic , erythroid series & Monocytes - POX Negative POX Reaction : POX Reaction Leukemia cells Leukemia cells Lymphocytic leukemia cells Sudan black B : Sudan black B Demonstrates lipoproteins and lipids Principle : Sudan black B (Fat soluble dye) Intracellular protein –bound lipids - Black intracytoplasmic granules - Diffuse black staining of cytoplasm Positive reaction Slide 64: Sudan black B positive - Pro , neutrophilic and meta myelocytes - Band cells - Segmented neutrophils Sudan black B negative - Lymphocytes - Megakaryocytes - Erythroblasts - Normoblasts - Platelets Neutrophil alkaline phosphatase (NAP) : Neutrophil alkaline phosphatase (NAP) Naphthol phosphate Naphthol Diazonium salt Insoluble precipitate NAP Bright Red color NAP present in mature neutrophils Other blood cells negative Chronic myeloid leukemia (NAP) : Chronic myeloid leukemia (NAP) Acid phosphatase (ACP) : Acid phosphatase (ACP) Principle : Specimen + Naphthol phosphate substrate Naphthol Diazonium salt Colored insoluble precipitate (red granules) (at the sites of acid phosphatase activity) Cells of hairy cell leukemia and auer bodies – Strongly positive Granulocytes - Positive RBC’s - Negative Slide 69: Hairy cell leukemia Auer bodies Naphthol AS-D chloroacetate esterase (AS-D NCE) : Naphthol AS-D chloroacetate esterase (AS-D NCE) Naphthol AS-D chloroacetate AS-D NCE H2O AS-D naphthol insoluble red ppt. + Corinth v salt Mast cells , granulocytes and their precursors (band & juvenile cells an promyelocytes) Positive α -Naphthol acetate esterase (α- NAE) : α -Naphthol acetate esterase (α- NAE) Principle similar to AS-D NCE α - Naphthyl acetate is used as a substrate and Pararosaniline as coupling azo dye Enzymatic activity results in intracytoplasmic dark red granules Periodic acid – schiff stain (PAS) : Periodic acid – schiff stain (PAS) Cytochemical stain (PAS) demonstrates Glycogen Glycogen Periodic acid Oxidation Aldehydes Parafuchsin(schiff reagent) Red color compound Slide 73: Stains demonstrating Iron (Gomori prussian blue stain) Common blood disorders : Common blood disorders Anemia Leukemia Myelodysplastic syndrome Multiple myeloma Primary ITP Anemia : Anemia Hyperplastic anemia Aplastic anemia Depending on BM study Hyperplastic anemia : Hyperplastic anemia Mainly seen in – Iron deficiency anemia Hemolytic anemia Hemorrhagic anemia Iron deficiency anemia : Iron deficiency anemia Hemolytic anemia : Hemolytic anemia Bone marrow Blood Aplastic anaemia(AA) [Acute] : Aplastic anaemia(AA) [Acute] Chronic Aplastic anaemia : Chronic Aplastic anaemia You do not have the permission to view this presentation. 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