finals - blood chem Flashcards by CHARLENE SAN JOSE

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

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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

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Peroxidase Stain/ Myeloperoxidase positive and negative in

Positive: Acute Myeloid Leukemia (AML)
Negative: Acute Lymphoid Leukemia (ALL)

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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

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Sudan Black B Stain positive and negative in

 Positive: Acute Myeloid Leukemia (AML)
 Negative: Acute Lymphoid Leukemia (ALL)

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differentiate granulocytic from monocytic leukemias

Stain for Esterases

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specific stain for esterase

Naphthol AS-D Chloroacetate Esterase (specific)

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non specific stain for esterase

α-naphthyl Acetate Esterase (non-specific)

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Naphthol AS-D Chloroacetate Esterase (specific)

positive and negative in

Positive: Granulocytic Leukemia
Negative: Monocytic Leukemia

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α-naphthyl Acetate Esterase (non-specific)

positive and negative in

 Positive: Monocytic Leukemia
 Negative: Granulocytic Leukemia

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gives a positive reaction to polysaccharides,
mucopolysaccharides, glycoproteins and glycogen

Periodic Acid Schiff (PAS) Reaction

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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

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Periodic Acid Schiff (PAS) Reaction positive and negative in

 Positive: Granulocytes, Platelet, Erythroleukemia/M6
 Negative: Acute Lymphoid Leukemia (ALL), Burkitt
Lymphoma

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Monoclonal or polyclonal antibodies against factor VIII-related
antigen have given positive results in megakaryoblastic leukemia

Factor VIII Antibodies

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ALL is positive and negativ in what stains

negative in MPO SBB NASDA Factor VIII

ANAE PAS positive and V

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differentiates CML from a leukemoid reaction seen in severe
infections

.Leukocyte Alkaline Phosphatase (LAP) Stain

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___ is used to differentiate
chronic myelogenous leukemia from leukemoid reaction

LAP (leukocyte alkaline phosphatase) grading

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Normal lap

20-100

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CML LAP scoring

<13

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Leukemoid Reaction lap scoring

> 100

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Polycythemia vera lap scoring

100-200

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Secondary polycythemia lap scoring

20-100

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0 grading in lap

No staining

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+ 1 grading in lap

Faint and diffuse staining

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+ 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 +)

2. 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

Megakaryocytic Leukemia is in what FAB classification

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

finals - blood chem Flashcards

(73 cards)