Chapter 7 - Cytochemistry Flashcards
is defined as the microscopic study and identification of chemical constituents within individual cells.
Cytochemistry
It is useful in the identification of malignant cell types on the basis of cytoplasmic or nuclear chemistry; cellular constituents that are present in abnormal form or amount; lack of cellular constituent; and cells exhibiting functional abnormalities
Cytochemistry
Enzymatic stains:
Non-Enzymatic Stains:
Stains NEUTROPHILS (the only leukocytes that contain this activity)
Leukocyte/Neutrophil Alkaline Phosphatase (LAP/NAP)
Differentiates Leukemoid reaction (LR) from Chronic myeloid leukemia (CML)
Leukocyte/Neutrophil Alkaline Phosphatase (LAP/NAP)
Leukocyte/Neutrophil Alkaline Phosphatase (LAP/NAP) Reference Value:
30 -185 LAP score
Inc LAP score:
- during the last trimester of pregnancy
- Polycythemia vera
- Aplastic anemia
- Hodgkin disease
- Multiple myeloma
- Obstructive jaundice
KAPLOWs Scoring (Count 100 cells and grade them as follows)
0 =
1+ =
2+ =
3+ =
4+ =
no staining
faint & diffuse staining
pale moderate amount of blue staining strong blue precipitate
deep blue or brilliant
Stain marker for primary granules & auer rods
Peroxidase/Myeloperoxidase (MPO)
Differentiates AML from ALL
Peroxidase/Myeloperoxidase (MPO)
(+) Activity: reddish-brown deposits (in cytoplasm of granulocytes and monocytes)
Peroxidase/Myeloperoxidase (MPO)
Note: (?) deteriorates; Stain should be done only on fresh specimens
Myeloperoxidase enzyme
For the diagnosis of Hairy cell leukemia (HCL)
Tartaric Acid Resistant Acid Phosphatase (TRAP)
Other Tartrate Resistant cells: Sezary cells; Histiocyte; T-cell of acute lymphocytic leukemia
Tartaric Acid Resistant Acid Phosphatase (TRAP)
Activity is indicated by purple to dark red granules in cytoplasm
Tartaric Acid Resistant Acid Phosphatase (TRAP)
For identification of Eosinophilic components
Cyanide-Resistant Peroxidase
(+) Activity: brown
Cyanide-Resistant Peroxidase
Marker for cells of Monocytic origin
a-Naphthyl Acetate Esterase & -Naphthyl Butyrate Esterase (Non-Specific Esterases/NSE)
Other cells (+) : Megakaryocytes, Platelets, Histiocytes, Plasmacytes, some T-lymphocytes
a-Naphthyl Acetate Esterase & -Naphthyl Butyrate Esterase (Non-Specific Esterases/NSE)
(+) Activity: red-brown/dark red
a-Naphthyl Acetate Esterase & -Naphthyl Butyrate Esterase (Non-Specific Esterases/NSE)
Marker for mature & immature Neutrophil and mast cells
Naphthol ASD Chloroacetate Esterase (Specific Esterase)
(+) Activity: bright red granules in cytoplasm
Naphthol ASD Chloroacetate Esterase (Specific Esterase)
This enzyme is stable and may last for months
Naphthol ASD Chloroacetate Esterase (Specific Esterase)
Stains DNA polymerase immunoperoxidase
Terminal Deoxyribonucleotidyl Transferase (TdT)
Marker for immature Lymphoid cell
Terminal Deoxyribonucleotidyl Transferase (TdT)
Positive in almost 90% cases of ALL
Terminal Deoxyribonucleotidyl Transferase (TdT)
Also useful in the detection of the “lymphoblastic transformation” of chronic myeloid leukemia (CML)
Terminal Deoxyribonucleotidyl Transferase (TdT)
Present in all hematopoietic cells and found in lysosomes
Acid Phosphatase
Activity is indicated by purple to red granules
Acid Phosphatase
Unstable, cannot be stored
Acid Phosphatase
Stains siderotic granules (for the diagnosis of sideroblastic anemia)
Prussian Blue stain
Used to detect hemosiderin in urine
Prussian Blue stain
Stain reacts w/ aldehyde groups in glycogen, mucoprotein, glycoproteins, and high molecular weight carbohydrates
Periodic Acid Schiff (PAS)
Positive in erythroblasts in Di Guglielmo disease / Erythroleukemia
Periodic Acid Schiff (PAS)
(-) Activity: bright fuchsia pink (Pattern of staining varies with each cell type)
Periodic Acid Schiff (PAS)
Also differentiates Acute Lymphoblastic leukemias
Periodic Acid Schiff (PAS)
Marker for phospholipids and lipids
Sudan Black B (SBB)
Gives the same information as Peroxidase in the interpretation
Sudan Black B (SBB)
Differentiates AML from ALL
Sudan Black B (SBB)
(+) Result: Dark purple-black granules
Sudan Black B (SBB)
Can be done on stored specimens.
Sudan Black B (SBB)
More sensitive than chloroacetate in the demonstration of mature & immature neutrophils and mast cells
Sudan Black B (SBB)
A metachromatic stain
Toluidine Blue O
Binds with mucopolysaccharides in blood cells
Toluidine Blue O
For the recognition of basophils and mast cells
Toluidine Blue O
(+) Result: Reddish-Violet
Toluidine Blue O