cytochemical stains
TRANSCRIPT
Cytochemical Stains Cutochemistry Identification of chemical constituent within the cell Celluler morphology is not changed Used in differentiation of acute leukemias 1. Enzymatic: myeloperoxidase, esterase, acid and alkaline phosphatase 2. Nonenzymatic: Sudan black B, periodic acid-Schiff, and terminal deoxynucleotidyl transferase
NAME Myeloperoxid ase
TYPE Enzyme
CONSITUENTS STAINED Marker for primary granules and Auer rods
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.
Sudan Black B (SBB)
Nonenzym e
Marker for phospholipids and lipids
Can be done on stored specimens Parallels myeloperoxid ase for interpretation Known as specific esterase Stable enzyme that lasts for months
Napthol AS-D Chloroacetat e Esterase
Enzyme
Marker of mature and immature neutrophils and mast cells
-Naphthyl Acetate Esterase - Naphtyl Butyrate Esterase
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
Neutrophil is the only leukocyte that contain this activity
Periodoc Acid-Schiff (PAS
Nonenzym e
Marker for glycoged, glycoproteins, mucoproteins, and high molecular weight carbohydrates
Terminal Deoxynucleot idyl Transferase (TdT)
Enzyme
DNA polymerase immunoperoxida se
Toluidine Blue
Nonenzym e
Binds with acid mucopolysaccha rides in blood cells
Strongly metachromati c
Useful for recognition of mast cells and basophils
Cytochemistry Flow Sheet in Acute Leukemia
BLAST CELL ON WRIGHTS STAIN
Peroxidase / Sudan Black
Acute nonlymphocytic leukemia M1 M2 M3 M4 M5 (+/-)
Acute Lymphocytic Leukemia
Esteras e
PAS
Naphthyl acetate Naphthyl Butyrate Positive MONOCYTIC M4 M5 M6
AS-D Chloroacetate Positive GRANULOCYTIC M1 M2 M3 M4 M5 (+/-)
Varaibl e L1 L2 M6
TDT L1 L2 L3
TRAP HAIRY CELL T-CELL
TDT = Terminal deoxytransaminase TRAP = Tartrate resistant acid phosphatase
Bone Marrow Analysis
Bone marrow aspiration
Erythroblasts % ANC
50% Erythroblast
Blasts % ANC
Blasts % NEC
>30% blasts