BM exam

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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

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