morphology of bone marrow aspirates -...
TRANSCRIPT
![Page 1: Morphology of bone marrow aspirates - inctrinctr-news.wdfiles.com/local--files/tutorials-in-hematopathology-in... · MORPHOLOGY OF BONE MARROW ASPIRATES Dr.Prasanna N Kumar Head](https://reader033.vdocuments.us/reader033/viewer/2022052710/5a9fe2707f8b9a6c178d59e6/html5/thumbnails/1.jpg)
MORPHOLOGY OF BONE MARROW ASPIRATES
Dr.Prasanna N Kumar Head – Department of
Pathology, Oman Medical College,
Oman
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BONE MARROW ASPIRATION
Sites
Sternum
Anterior or posterior iliac spines
Aspiration from multiple sites may be needed
eg: in aplastic anaemia – partial involvement
of the bone marrow
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Do not aspirate more than 0.3 ml of marrow fluid from a single site for morphological examination as this increases peripheral blood dilution.
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APPROACH TO A MAROW
ASPIRATE
Macroscopic evaluation –
check for presence of particles
Check that there is no dilution
with peripheral blood- leads to
an erroneous increase in
lymphocytes and neutrophils
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ESTIMATION OF CELLULARITY
Critical information – age of patient
A young child – about 80% of intertrabecular space
occupied by haemopoietic cells
A 75-year-old the average has about 30%
Semiquantitative assessments of cellularity –
hypoplasia, within normal limits, hyperplasia
Better assessment with biopsy
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COMPARING NORMO, HYPER, & HYPOCELLULAR MARROWS
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LOW POWER – A SNEAK PREVIEW OF THE MARROW!
Look at:
Cellularity of the marrow
Identify megakaryocytes
Look for aggregates of abnormal cells – eg:
granulomas, metastatic deposits
Identify macrohages
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BONE MARROW GRANULOMAS
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LOW POWER
Films without fragments are worth examining
However, assessment of cellularity and
megakaryocyte numbers is unreliable and dilution
with peripheral blood may lead to lymphocytes and
neutrophils being over-represented in the
differential count.
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ARTEFACTS
Process of slide preparation - bare nuclei. D/D – monotonous cells of malignancy
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BONE MARROW AT HIGHER
POWERS
Higher power (x40, x100)
Identify all stages of maturation of myeloid and erythroid cells.
Determine the M:E ratio
Perform a differential count
Look for areas of BM necrosis.
Assess the iron content.
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M:E RATIO
Expression of relative proportions of myeloid (granulocytic and monocytic cells)to erythroid cells
Rough comparative index – exclude eosinophils,
basophils, lymphocytes, plasma cells
Must comment on lymphocytes and plasma cells
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Decreased M:E ratio (1:1) - increased erythroid precursors
Ratio of limited use if both series are equally
depressed or stimulated (normal M:E)
Significant if ratio is off by >2X= eg >5:1 or less than 1.2:1
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MARROW DIFFERENTIAL COUNT
Not routinely necessary but an excellent practice –
good for a beginner to familiarise with the marrow
cells
Study morphology in fields that are evenly dispersed
– scan many fields under LP and then oil
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MARROW DIFFERENTIAL COUNT
Count atleast 200 cells and also note the
smudge/unidentifiable cells
Any borderline abnormality - e.g. in the
number of blasts, lymphocytes or plasma
cells, perform a 500 cell differential count
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EVOLUTION OF HAEMATOPOIETIC
CELLS
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ERYTHROPOIESIS
& MYELOPOIESIS
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PROERYTHROBLAST
Large cell, moderately to strongly basophilcic cytoplasm, round nucleus, finely stippled chromatin pattern. Nucleoli are sometimes apparent.
There may be a paler staining area of cytoplasm surrounding the nucleus.
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ERYTHROID SERIES
Early (basophilic) erythroblast - similar to a proerythroblast but
smaller without visible nucleoli
Intermediate (polychromatophilic)
erythroblast and late erythroblast
(orthochromatic) – cell
size reduction, reduction in
cytoplasmic basophilia and
increase in chromatin clumping.
Cytoplasm of the late
erythroblast - pink tinge due to
haemoglobin.
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MULTINUCLEAR
ERYTHROBLAST
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NORMAL GRANULOCYTE
PRECURSORS IN BONE MARROW
Myeloblast - high
nucleocytoplasmic ratio,
diffuse chromatin pattern
and nucleolus.
Promyelocyte – larger,
lower nucleocytoplasmic
ratio, abundant azurophilic
granules.
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MYELOCYTES &
METAMYELOCYTES
Myelocytes - smaller than
promyelocytes, contain specific granules that indicate whether they are of neutrophil, eosinophil or basophil lineage.
The nucleolus is no longer visible.
Some indentation of the nucleus in metamyelocytes
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METAMYELOCYTES
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BAND OR JUVENILE
NEUTROPHILS Of neutrophil lineage with
non-segmented nuclei.
Band forms, stab forms
Less mature than
segmented neutrophils.
.
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TIME LINE
Maturation time from proerythroblast to reticulocyte -
5 days
Maturation from myeloblast to neutrophil - 7 days
At times of need the days are shortened.
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MEGAKARYOCYTES
< 0.1% of all marrow cells
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NONHAEMATOPOIETIC CELLS OF THE NORMAL MARROW
OSTEOCLASTS
OSTEOBLASTS
MAST CELLS STAINED WITH TOLUIDINE BLUE
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BM IRON STORES
Storage iron - stains blue, assessed in bone marrow fragments.
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ERYTHROID HYPERPLASIA
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GRANULOCYTIC HYPERPLASIA
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MEGALOBLASTIC ANAEMIA Patient with anaemia, hyperpigmentation of knuckles, glossitis Hb – 8g/dL MCV – 110 fl TWBC – 2.5 X 109/l PLT – 75 X 109/l
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ACUTE LEUKAEMIA
23-year-old male with lymphadenopathy and hepatosplenomegaly
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MULTIPLE MYELOMA
66-year-old male with severe back pain, anaemic, ESR- 150 mm/hour PS - rouleaux
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11-month-old baby with anaemia and hepatosplenomegaly NEIMANN-PICK DISEASE – FOAMY MACROPHAGES
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CASE - 7 1 ½ month female child with fever, abdominal distension,
oliguria, altered sensorium, jaundice +, bilateral
peripheral lymphadenopathy +, hepatosplenomegaly +
Another sibling had the same problem and succumbed
Familial haemophagocytic
lymphohistiocytosis
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CASE - 6
A 16-year-old apparently healthy school boy
comes for evaluation of his hepatosplenomegaly
and hypercholesterolaemia
He had been diagnosed as having Niemann-Pick
disease when he was 5 years old at another
hospital
Serum cholesterol 154 mg/dl, triglycerides 227 mg/dl
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SEA BLUE HISTIOCYTES