Leukocyte Alkaline Phosphatase
EMS Catalog #26913
Sample
Capillary Blood Or Heparinized Venous Blood Films
Fixative
Procedure
- 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.
Results
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.
Scoring Criteria
| 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.
Normals
Adults - Leukocyte Alkaline Phosphatase Score Of 13 To 130.
References
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.
Online Ordering
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.
