BM exam

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Presentation Transcript

Slide 1: 


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: 


Normoblast Basophilic normoblast : 

Normoblast Basophilic normoblast

polychromatic normoblast ortho-chromatic normoblast : 

polychromatic normoblast ortho-chromatic normoblast

Net erythrocyte Erythrocyte : 

Net erythrocyte Erythrocyte

Slide 16: 


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: 


Lymphoblast : 

Lymphoblast Prolymphocyte Lymphocyte

Slide 26: 


Monoblast : 

Monoblast Promonocyte Monocyte

Slide 28: 


Plasmablast : 

Plasmablast proplasmacyte plasmacyte

Slide 30: 


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 : 


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 •Pelvis-40% •Vertebrae-28% •Cranium-mandible-13% •Ribs-8% •Sternum-2% •Ends of long bones-8%


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 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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