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BONE MARROW PUNCTURE ( BMP )
Prof. Adi Koesoema Aman SpPK(KH)
Divisi Hematologi Departement Patologi
KLinik FK USU / RSUP.H.A.Malik ,Medan
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BONE MARROW PUNCTURE ( BMP ) .
1. Aspirasi .- Red Maroow .
2. Aspirasi & Biapsi .
- Structure Tulang .
- Trabeculae Tulang .
- Sinusoid .
- Jaringan hematopoitik dan
non-hematopoitik
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1 . Aspirasi :
- Jarum Salah .
2. Aspirasi Biopsi :
- Jarum Silvermman .
- Islamic Needle .
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Bone Marrow -Definition
Bone refers to location ofbone marrow after birth
Marrow: Latin for middle.
Hence Bone Marrow is located
in middle of bone.
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DISTRIBUTION OF BONE MARROW
Pelvis-40%Vertebrae-28%
Cranium-mandible-13%
Ribs-8%Sternum-2%
Ends of long bones-8%
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TYPES OF MARROW
Yellow marrow (inactive)
composed primarily of fat
Red marrow (active in
hematopoiesis)
M t dl
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Marrow puncture needle
- Salah
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X- sternal aspiration sites Tibial aspiration site
Posterior iliac crest aspiration
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BM SITES AND INDICATIONS
FOR ASPIRATIONS
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 for
aspiration)
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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
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Normal cellularity
Hypocellular Hypercellular
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Staining of cells
Identification of various types of
blood cells
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CYTOLOGICAL
EXAMINATION OF
BONEMARROW
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Myeloblast Promyelocyte
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MetamyelocyteNeutrophil ic myelocyte
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Band granulocyte segmented granulocyte
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Band granulocyte segmented granulocyte
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Leukemia cells
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LYMPHOCYTE
SERIES
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Lymphoblast Prolymphocyte Lymphocyte
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Lymphocyticleukemia cells
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MONOCYTE SERIES
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MonoblastPromonocyte Monocyte
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PLASMA SERIES
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Plasmablastproplasmacyte plasmacyte
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MEGAKARYOCYTE
SERIES
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Megakaryoblast promegakaryocyte
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Granular megakaryocyte Thrombocytogenous megarkaryocyte
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Other
cells
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Mesh phagocytes Fat cells
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Endothelial cel ls Osteoblasts
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Osteoclasts
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PENGENALAN SEL HEMATOPOITIK
UNTUK MENGENALI JENIS SEL DARAHDAPAT DILAKUKAN DGN CARA :
1. MORFOLOGI SEL .
1.1. PENGECATAN RUTIN .
1.2. PENGECATAN SITO-KIMIAWI .
2. PENENTUAN CLUSTER DIFFERENTIATION( CD ) FLOWCYTOMETRI IMMUNO
PHENOTYPING
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1.1. PENGECATAN RUTIN .
PENGECATAN RUTIN DILAKUKAN DG CARA :
1. MGG ( MAYGRUNWALDGIEMSA
STAIN .
2. ROMANOWSKY STAIN .
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1.2. PENGECATAN SITO-KIMIAWI
PENGECATAN SITO - KIMIA DIGUNAKAN
UNTUK DIAGNOSE BANDING DAN
KLASIFIKASI LEUKEMIA , YAITU UNTUK :
1. MEMBEDAKAN MIELOBLAST DAN
MONOBLAST .
2. MENENTUKAN SEL LIMFOBLAST .
3. MEMBEDAKAN LEKOSITOSIS DGNREAKSI LEKEMOID .
4. MEMPELAJARI KELAINAN ATAU
DEFISIENSI ENZYM DLM NEUTROFIL
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Cyto - Staining methods
POX-NAENAPPASFe dyes
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(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
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POX Reaction
Leukemia cellsLeukemia cells Lymphocyticleukemia cells
Sudan black B
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Sudan black B
Demonstrates lipoproteins and
lipidsPrinciple :
Sudan black B (Fat soluble dye)
Intracellular proteinbound lipids
-Black intracytoplasmic granules
- Diffuse black staining of cytoplasm
Positive reaction
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Sudan black B positive
- Pro , neutrophilic and meta myelocytes
- Band cells
- Segmented neutrophils
Sudan black B negative- Lymphocytes- Megakaryocytes
- Erythroblasts
- Normoblasts
- Platelets
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Neutrophil alkaline phosphatase (NAP)
Naphthol phosphateNaphthol
Diazonium salt
Insoluble precipitate
NAP
Bright Red color
NAP present in mature neutroph i ls
Other blood cells negat ive
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Chronic myeloid leukemia (NAP)
A id h h t (ACP)
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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 hair y cell leukemia and auer bodiesStrongly positive
Granulocytes - Positive
RBCs- Negative
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Hairy cell leukemia Auer bodies
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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
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-Naphthol acetate esterase (- NAE)
Principle similar to AS-D NCE
- Naphthyl acetate is used as a
substrate andPararosaniline as coupling azo dye
Enzymatic activity results in
intracytoplasmic dark red granules
P i di id hiff t i (PAS)
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Periodic acid schiff stain (PAS)
Cytochemical stain (PAS) demonstrates
Glycogen
Glycogen Periodic acid
Oxidation
Aldehydes
Parafuchsin(schiff reagent)
Red color compound
Stains demonstrating Iron
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Stains demonstrating Iron
(Gomori prussian blue stain)
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Common blood disorders
Anemia
Leukemia
Myelodysplastic syndrome
Multiple myeloma
Primary ITP
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MA KASIH IYA SAYA DI SURAMADU SURABAYA