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of acute leukemias 1. Enzymatic: myeloperoxidase, esterase, acid and alkaline phosphatase 2. Nonenzymatic: Sudan black B, periodic acid-Schiff, and terminal deoxynucleotidyl transferase
TYPE Enzyme
INTERPRETAT ION Peroxidase activity produces dark brown granules in cytoplasm of granulocytes and monocytes. Neutrophil and precursors Monocyte weak positive Dark purpleblack granules in neutrophil precursors Lymphoblasts are negative Enzyme activity results in bright red granules in cytoplasm of neutrophils,
IMPORANT INFORMATIO N Myeloperoxid ase enzyme deteriorates. Stain should be done only on fresh specimens.
Nonenzym e
Can be done on stored specimens Parallels myeloperoxid ase for interpretation Known as specific esterase Stable enzyme that lasts for months
Enzyme
Enzyme
Enzyme
Marker for monocytes, megakaryocytes, and plasma cells Identifying monocytes, promonocytes, and monoblasts Present in all hematopoietic cells and found in lysosomes Marker for hairy cell leukemia
neutrophil precursor, and mast cells. Monocytes stained brown Enzyme activity results in dark, red precin cytoplasm Activity is indicated by purple to red granules Activity is indicated by purple to dark red granules in cytoplasm 100 cell count is done. Neutrophils are scored from 0 with no activity to 4 with large amount of activity. Activity results in bright fuchsia pink. Pattern of staining varies with each cell type. Is present in 90% cases of ALL
Monocytes are weakly potive. Known as nonspecific esterase (NSE) Known as nonspecific esterase (NSE) Cannot be stored Excellent marker for hairy cell leukemia Used to differentiate CML form leukemoid reaction. CML has decreased activity. Lymphoblasti c leukemias show blockly or crunchy pattern. L1 and L2, block pattern L3 negative Erythroblasts in M6 Leukemia also positive Used to differentiate AML from ALL
Acid Phosphatase Tartrate Resistant Acid Phosphatase Leukocyte Alkaline Phosphatase (LAP)
Enzyme
Enzyme
Enzyme
Nonenzym e
Marker for glycoged, glycoproteins, mucoproteins, and high molecular weight carbohydrates
Enzyme
Toluidine Blue
Nonenzym e
Strongly metachromati c
Esteras e
PAS
Varaibl e L1 L2 M6
TDT L1 L2 L3
Erythroblasts % ANC
<50% Erythroblast
>50% Erythroblast
Blasts % ANC
Blasts % NEC
>30% blasts
<30% blasts
<30% blasts
>30% blasts
AML
MDS
MDS
M6
M1 M5