Dr. P.G.

Subramanian
Assistan Professor and Officer incharge Hematopathology
laboratory
Tata Memorial Hospital
Diagnostic
Thrombocytopenia
Bicytopenia
Pancytopenia
Unexplained organomegaly/ Fever/anemia
Suspected acute leukemia
Staging for lymphomas
Baseline evaluation
Chronic myeloproliferative disorders
Myeloproliferative myelodysplastic disorders
Plasma cell dyscrasias
Post treatment evaluation
Post induction, Day 10 marrows
Clinical suspicion of relapse
Unexplained cytopenias
Unexplained fever
Cytochemistry
Perls iron stain
Myeloperoxidase
Sudan black B
Non specific esterases ( -naphthyl acetate or -
naphthyl butyrate esterase)
Immunophenotyping by flow cytometry
Cytogenetics
Molecular testing
Culture
Normal bone marrow
Megakarocyte
Increased blasts greater than 20%.
Acute leukemia/ CML blast crisis (presence of
a documented chronic phase)/ rarely high
grade non hodgkins lymphoma
Various morphology of blasts
Cytochemistry/ Immunophenotyping
/Cytogenetics/ molecular studies
Myeloperoxidase- If positive Acute Myeloid
Leukemia. If negative then
immunophenotyping is essential.
Nonspecific esterase positive in Acute
Monoblastic and Acute Monocytic leukemias
Essential for diagnosis of
Acute Lymphoblastic Leukemia (B cell and T cell
type)
Acute Myeloid Leukemia M0
Acute Monoblastic Leukemia M5
Acute Megakaryoblastic Leukemia M7
2 techniques
Immunohistochemistry
Flow cytometry using
immunofluorescence
Comparison of the two methods currently
Flow cytometry
(Immunofluorescence)
Immunohistochemistry
Multiparametric evaluation of
single cells
Higher sensitivity
Requires significant
concentration of population of
interest in the sample
Gives accurate counts and
percentages
Fixation and antigen retrieval
problems
Morphological confirmation of
population of interest
Can detect a few cells in a
section
Certain degree of subjectivity in
interpretation of weak positivity
Essential for diagnosis of
Philedelphia chromosome positive leukemias and
CML blast crisis
Acute promyelocytic leukemia
AML
AML-M3
Case: APML
24/06/2010
8 yr male child referred from outside as case of Acute
Lymphoblastic Leukemia.
Thrombocytopenia
Bone marrow done dilute with scattered 40% blasts
like cells and 60% lymphocytes
MPO - Negative
FCM - Outside diagnosed as CALLA positive ALL
with CD19+, CD10+, HLADR+ & small percentage
of CD34+ cells
R1
Hematogones
Hematogones
Another case : new case
4 yrs female child,
High WBC count, thrombocytopenia and low Hb
BM 90% blasts, MPO negative
Immunophenotyping done
R1
HEMATOGONES CALLA+ ALL BLASTS
12 yr old boy with hepatosplenomegaly and
lymphadenopathy with complaints of
weakness and difficulty in breathing.
Morphology
Diagnosis:
T- cell Acute Lymphoblastic Leukemia
5 year old Male child Presented with weakness and severe Pallor. Hb 4 gm
/ dL
Given one unit of blood at the local clinic
CBC showed few blasts and referred.
Bone marrow on examination shows 45% blasts mixed with lymphocytes
and normal myeloid series cells
Diagnosis????
Dyserythropoiesis
MDS
43 Yr Male
Hairy Cell
Leukemia
HCL
Reed Stern berg Cell