ENZYMATIC STAIN

is a normal constituent of the primary granules of the myelocytic cells

Myeloperoxidase

Myeloperoxidase is observed as early as

promyelocyte

PEROXIDASE/MYELOPEROXIDASE (MPO) is Stain marker for

primary granules

auer rods (fused primary granules)

immature myeloid cells

PEROXIDASE/MYELOPEROXIDASE (MPO) Differentiates

AML from ALL

PEROXIDASE/MYELOPEROXIDASE (MPO) stain positive for

AML

granulocyte

PEROXIDASE/MYELOPEROXIDASE (MPO) stain negative for

ALL

lymphocytes

PEROXIDASE/MYELOPEROXIDASE (MPO) positive result color

reddish-brown deposits

Used to detect the granulocyte of monocytic origin

ALPHA-NAPHTHYL ACETATE ESTERASE & ALPHANAPHTHYL

BUTYRATE ESTERASE (NON-SPECIFIC ESTERASES/NSE)

Other positive cell for ALPHA-NAPHTHYL ACETATE ESTERASE & ALPHANAPHTHYL BUTYRATE ESTERASE (NON-SPECIFIC ESTERASES/NSE)

Megakaryocytes,

Platelets,

Histiocytes,

Plasmacytes,

some T-lymphocytes

ALPHA-NAPHTHYL ACETATE ESTERASE & ALPHANAPHTHYL BUTYRATE ESTERASE (NON-SPECIFIC ESTERASES/NSE) Positive Activity:

red-brown/dark red

Principle: Differential count involving neutrophils

LEUKOCYTE/NEUTROPHIL ALKALINE PHOSPHATASE (LAP/NAP)

LEUKOCYTE/NEUTROPHIL ALKALINE PHOSPHATASE (LAP/NAP)

Differentiates

Leukemoid reaction (LR) from

Chronic myeloid leukemia (CML)

LAP Reference Value:

30 –185 LAP score

Increase LAP score is observed in the following:

o during the last trimester of pregnancy - Hodgkin disease

o Polycythemia vera - Multiple myeloma

o Aplastic anemia - Obstructive jaundice

Normal to High LAP score

: Leukemoid reaction

Below than normal value to zero LAP score

CML

For the diagnosis of Hairy cell leukemia (HCL)

TARTARIC ACID RESISTANT ACID PHOSPHATASE (TRAP)

ACP is Detected in almost all blood cells but when treated with tartaric acid it is inhibited except

isoenzyme number 5

TARTARIC ACID RESISTANT ACID PHOSPHATASE (TRAP) should be fixed and stored at

20°C and can be used atleast 2 weeks

Other Tartrate Resistant cells:

Sezary cells;

Histiocyte;

T-cell of acute lymphocytic leukemia

TRAP Activity is indicated by

purple to dark red granules in cytoplasm

For identification of Eosinophilic components

CYANIDE-RESISTANT PEROXIDASE

CYANIDE-RESISTANT PEROXIDASE Positive Activity:

brown

Marker for mature & immature Neutrophil and mast cells.

NAPHTHOL ASD CHLOROACETATE ESTERASE (SPECIFIC ESTERASE)

NAPHTHOL ASD CHLOROACETATE ESTERASE (SPECIFIC ESTERASE) Substrate is the

chloroacetate

NAPHTHOL ASD CHLOROACETATE ESTERASE (SPECIFIC ESTERASE)

Is Use to identify

immature or primitive granulocyte ALM1-ALM3

NAPHTHOL ASD CHLOROACETATE ESTERASE (SPECIFIC ESTERASE)

Positive Activity:

bright red granules in cytoplasm

This enzyme is stable and may last for months

NAPHTHOL ASD CHLOROACETATE ESTERASE (SPECIFIC ESTERASE)

Stains DNA polymerase immunoperoxidase

TERMINAL DEOXYRIBONUCLEOTIDYL TRANSFERASE (TDT)

TERMINAL DEOXYRIBONUCLEOTIDYL TRANSFERASE (TDT) is a Marker for

immature Lymphoid cell

TDT is Positive in almost 90% cases of

ALL

TDT is also useful in the detection of the

“lymphoblastic transformation” of chronic myeloid leukemia (CML)

more than or equal to 30% blast cells

Acute leukemia

Present in all hematopoietic cells and found in lysosomes

ACID PHOSPHATASE

 Activity is indicated by purple to red granules

 Unstable, cannot be stored.

ACID PHOSPHATASE