Stain Color Positive Reaction Positive cell Negative cell

1.Myeloperoxidase – DARK BROWN Neutrophils, eosinophils Lymphoid cells, Diamino benzidine + H2O2
identifies Auer rods in Weak: monocytes megakaryocytes & →peroxidase → H2O +
AML erythroid cells colored precipitate ( DARK
-diffusely brown RBCs: BROWN)
pseudoperoxidase
- primary granules & auer
rods

2. Cyanide resistant- BROWN Eosinophils

peroxidase stain -eosinophilic leukemia
• sodium cyanide
• inhibit peroxidase other
than those found in
eosinophils
3. Esterases a. Specific esterase or Naphthol AS-D a. Neutrophils, mast cell a. Eosinophils,
-hydrolyses Chloroacetate esterase monocytes,
-marker for mature & immature neutrophils & mast lymphocytes
cells
-enzymatic hydrolysis of ester linkages occur
BRIGHT RED GRANULES
b. Alpha-naphthyl acetate esterase
c. Granulocytes,
-incubation with alpha-naphthol at an alkaline Ph megakaryocytes,
- sodium fluoride: inhibits the monocytic enzyme lymphocytes
RED BROWN

c. Alpha –naphthyl butyrate esterase b. Monocytes, histiocytes,

DARK RED megakaryocytes, plasma cells, some
T lymphocytes (clot like pattern)
 d. Combination of both Alpha –naphthyl butyrate-
Chloroacetate esterase
- monocytes & histiocytes
DARK RED PRECIPITATE
-neutrophils
BLUE GRANULES

c. Monoblasts, Promonocytes,
Monocytes
-AMML, AMoL, CMML

4. Leukocyte Alkaline BLACK BROWN GRANULES To calculate the LAP Score:

Phosphatase (LAP) 1. the # of cells is multiplied by
Scoring: the grade & the products are
0 – no granules added together
1+ - faint, few granules 2. RR; 30-100
(<50%) ↓LAP activity
2+ - moderate (50-75%) 1. CGL
3+ - strong, numerous 2. PNH
(75%)
4+ - intense, cytoplasm is 3. SA
packed with granules 4. Eosinophilia
(100%) ↑ LAP activity
1. LR
2. CNL
3. Infection
4. PCCU
5. Third trim of pregnancy
6. CGL Blast crisis or with
infection

5. Periodic Acid Schiff MAGENTA Neutrophils, basophils, eosinophils, Lymphocytes,

(PAS) megakaryocytes, platelets, eyrthrocytes
-stains glycogen, monocytes
polysaccharides,
monosaccharides,
glycoproteins

6. Sudan Black B (SBB) BROWN BLACK Auer rods Lymphocytes

-diazo dye that stain
phospholipids, neutral
fats & sterols

7. Acid Phosphatase PURPLISH TO DARK RED GRANULES Lysosomes in all hematopoietic cells
8. Tartrate resistant PURPLISH TO DARK RED GRANULES
acid phosphatase HCL ISOENZYME 5
(TRAP)
 9. Terminal
deoxynucleotidyl
transferase (TdT)

10. Toluidine Blue RED TO PURPLE METACHROMATIC Basophils, mast cells

-reacts with acid GRANULES
mucopolysaccharides -excessive basophils: mastocytosis

Cytochemical Staining