Leukocyte Alkaline Phosphatase
EMS Catalog #26913
Capillary Blood Or Heparinized Venous Blood Films
- Fix blood films in Buffered Acetone Fixative for 30 seconds.
- Wash Blood films in running tap water for 30-60 seconds. Air dry or blot.
- Stain films for exactly 10 minutes in staining Solution. *To prepare staining solution: Dissolve 5mg of Naphthol AS-BI Phosphate in 0.2 to 0.3ml of Dimethyl Formamide. Add 60m of Propanediol Buffer. Add 40mg Fast Red Violet Salt LB or Fast Violet B Salt. Filter and use at once.
- Wash blood films in running tap water for 30 to 60 seconds.
- Counterstain in Hematoxylin Solution filtered just prior to use, for 5 - 8 minutes.
- Rinse in running tap water 1 to 2 minutes. Air dry or blot carefully.
- Examine microscopically under an oil immersion lens.
Intensity of precipitated dye in the cytoplasm of neutrophilic leukocytes varies. One hundred are scored from 0 to 4+ based on the intensity of the precipitated dye.
|CELL RATING||AMOUNT **%||Ppt'd azo dye In cytoplasm Size||INTENSITY||BACKGROUND OF CYTOPLASM|
|1+||<50||Small||Faint to moderate||Colorless to very pale pink|
|2+||50-80||Small to Med.||Moderate to strong||Colorless to pale pink|
|3+||80-100||Med. To Large||Strong||Colorless to pink|
|4+||100||Med. To Large||Brilliant||Not visible|
* When there is no precipitate azo dye in the cytoplasm and the background is not stained, the cells are read as zero.
** Percentage of volume of cytoplasm occupied by azo dye precipitate.
Adults - Leukocyte Alkaline Phosphatase Score Of 13 To 130.
Williams, W.J., Beutler, El, Erslev, A.J. and Rundles R.W., Hematolog y, 2 nd ed., McGraw-hill , NY , C. 1977, p1628-1629.
Kaplow, L.S., AM.J., Clin. Patbol ., 39:439, 1963.
Kaplow, L.S. and Burstone, M.S., Nature , 200:690, 1963.
Buffered Acetone Fixative, Naphthol AS-BI Phosphate, Dimethyl Formamide, Fast Red Violet Salt LB, or Fast Violet B Salts, Propanediol Buffer and Hematoxylin Counter Stain for Leukocyte Alkaline Phosphatase are available online from the EMS Catalog. For ordering or product information, click here.