finals - blood chem Flashcards
Uses of Cytochemical Stains
- Used in the study of cell differentiation and in the classification of
acute leukemias - Used in the differentiation of leukocytosis and leukemoid reactions
from genuine myeloproliferative disorders - Used in the characterization of lymphoproliferative disorders
- Used in the demonstration of free iron, Hb derivatives, DNA, RNA,
and red cell enzymes in RBC and WBC
In the presence of hydrogen peroxide, MPO in granulocytes
oxidizes benzidine dihydrochloride from a colorless to a reddish
brown derivative at the site of the enzyme
Peroxidase Stain/ Myeloperoxidase
Peroxidase Stain/ Myeloperoxidase positive and negative in
Positive: Acute Myeloid Leukemia (AML)
Negative: Acute Lymphoid Leukemia (ALL)
It stains lipids, such as sterols, neutral fats, and phospholipids
because of the solubility of the dye in lipid particles and appears
dense black or bluish black
Sudan Black B Stain
Sudan Black B Stain positive and negative in
Positive: Acute Myeloid Leukemia (AML)
Negative: Acute Lymphoid Leukemia (ALL)
differentiate granulocytic from monocytic leukemias
Stain for Esterases
specific stain for esterase
Naphthol AS-D Chloroacetate Esterase (specific)
non specific stain for esterase
α-naphthyl Acetate Esterase (non-specific)
Naphthol AS-D Chloroacetate Esterase (specific)
positive and negative in
Positive: Granulocytic Leukemia
Negative: Monocytic Leukemia
α-naphthyl Acetate Esterase (non-specific)
positive and negative in
Positive: Monocytic Leukemia
Negative: Granulocytic Leukemia
gives a positive reaction to polysaccharides,
mucopolysaccharides, glycoproteins and glycogen
Periodic Acid Schiff (PAS) Reaction
Periodic acid is an oxidizing agent that converts hydroxyl groups
on adjacent carbon atoms to aldehydes and further combined with
Schiff’s reagent to give a red color product
Periodic Acid Schiff (PAS) Reaction
Periodic Acid Schiff (PAS) Reaction positive and negative in
Positive: Granulocytes, Platelet, Erythroleukemia/M6
Negative: Acute Lymphoid Leukemia (ALL), Burkitt
Lymphoma
Monoclonal or polyclonal antibodies against factor VIII-related
antigen have given positive results in megakaryoblastic leukemia
Factor VIII Antibodies
ALL is positive and negativ in what stains
negative in MPO SBB NASDA Factor VIII
ANAE PAS positive and V
differentiates CML from a leukemoid reaction seen in severe
infections
.Leukocyte Alkaline Phosphatase (LAP) Stain
___ is used to differentiate
chronic myelogenous leukemia from leukemoid reaction
LAP (leukocyte alkaline phosphatase) grading
Normal lap
20-100
CML LAP scoring
<13
Leukemoid Reaction lap scoring
> 100
Polycythemia vera lap scoring
100-200
Secondary polycythemia lap scoring
20-100
0 grading in lap
No staining
+ 1 grading in lap
Faint and diffuse staining
+ 2 grading in lap
Pale with moderate amount of
blue staining
+3 grading in lap
Strong blue precipitated
staining
+4 grading in lap
Deep blue or brilliant staining
with no visible cytoplasm
diagnostic tool for confirmation of hairy cell leukemia
Acid Phosphatase (Tartrate Resistant)
- stain for iron - it gives a positive reaction in cases of cells
containing iron, ringed sideroblasts, and siderocytes
Prussian Blue/ Perl’s Stain
differentiates basophilic leukemia and mast cell leukemia from
other diseases
positive result is metachromatic granules which are colored
reddish violet and is observed in mast cells and basophilic
leukemia
Toluidine Blue
Peroxidase test for
Myeloperoxidase
(MPO)
Peroxidase indication
AML AMMoL (weak +)
- Sudan Black B test for
lipid
Specific Esterase test or
Granulocyte
precursor
non-specific
Esterase test for
Monocyte
precursor
PAS (Periodic
Acid Schiff) test for
glycogen
Tartrate Resistant
Acid Phosphatase
(TRAP) test for
hairy Cell Leukemia
NBT (Nitroblue
tetrazolium test) test for
Granulocyte
function
TdT (Terminal
deoxynucleotidyl
transferase)
test for
Early T and B
lymphocyte
LAP (leukocyte
alkaline
phosphatase) test for
leukomoid reaction and leukemia
Prussian Blue test for
irron
Phyloxine test for
acid granules
Acute lymphocytic leukemia (ALL)
L1 Small lymphoblasts
o Most common childhood leukemia
o Found in young adult
o Homogenous appearance
o ? prognosis
Acute lymphocytic leukemia (ALL)
L2 - Small and large lymphoblasts
o Most common in adults
o Heterogenous appearance
L3 Large homogeneous lymphoblasts
o Leukemic phase of Burkitt lymphoma
differentiates basophilic leukemia and mast cell leukemia from other diseases
Toluidine Blue
positive result is metachromatic granules which are colored reddish violet and is observed in mast cells and basophilic leukemia
Toluidine Blue
Childhood ALL percentages
L1 – 70% predominantly
L2 - 27%
L3 – 3% or less - Burkitt cell type
Adult ALL percentage
L1 – 30%
L2- 65%
L3 – 5%
Acute myeloid leukemia (AML)
M1 - Myeloblasts without maturation
o Show 90% or more marrow myeloblast
o Presence of the auer rods
Acute myeloid leukemia (AML
M2- Myeloblasts with maturation (best AML prognosis)
Show less than 90% marrow myeloblast
acute myeloid leukemia
M3-
Hypergranular promyelocytic leukemia
acute myeloid leukemia
M3V Variant,
microgranular promyelocytic leukemia
acute myeloid leukemia
M4- Myeloblast and Monoblast
o More or less 20% best on WHO if FAB greater than 30% marrow
o in proliferation of unipotenial stem cell Accounts 30% of AML
acute myeloid
M5
monoblast
Poorly differentiated monocytic leukemia
M5- Monoblast
Well differentiated monocytic leukemia
M5- Monoblast
M5- Monoblast
Characterized by greater than, less than or equal____ based on WHO
20%
M5 - monoblast
IN FAB precipitation greater than ___% marrow
monoblast
30%
M5 - monoblast is ___% AML
10%
Variant of M5 monoblast
___
Seen in children with 80%
monoblast
M5a
Variant of M5 monoblast
___
Seen in middle age adult with less
than 80% monoblast
M5b
Erythroleukemia/DiGuglielmo syndrome is in what fab classification
M6
Megakaryocytic Leukemia is in what FAB classification
m7
pleomorphic undifferentiated cells with cytoplasmic
blebs
M7- Megakaryocytic Leukemia
myelofibrosis or ↑ BM reticulin
M7- Megakaryocytic Leukemia
positive for platelet peroxidase antifactor VIII
M7- Megakaryocytic Leukemia
proliferation of megakaryoblast and atypical
megakaryocytes in the bone marrow
M7- Megakaryocytic Leukemia
less than 1% AML
M7- Megakaryocytic Leukemia
marrow aspiration result in dry top, blood shows
pancytopenia this is difficult to diagnose with
cytochemical stain
M7- Megakaryocytic Leukemia
rarest type of AML
M7- Megakaryocytic Leukemia
-Blood shows pancytopenia, this is difficult to diagnose
with cytochemical stains
M7- Megakaryocytic Leukemia
