Pk 5-Kuliah Mhs Fkg Usu Bmpkbk

7/28/2019 Pk 5-Kuliah Mhs Fkg Usu Bmpkbk

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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
http://www.clinet.com.cn/clinetforum/cgi-bin/attachment.cgi?forum=18&topic=73&type=.jpg&name=1_1097667477http://www.clinet.com.cn/clinetforum/cgi-bin/attachment.cgi?forum=18&topic=73&type=.jpg&name=1_1097667477


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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

Pk 5-Kuliah Mhs Fkg Usu Bmpkbk

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