A stain for neutrophils.

Intended Use :

NEUTROCOLOR offers a rapid means for detection of neutrophils in samples of blood and bone marrow, and compliments the more complex monoclonal antibody latex sphere and myeloperoxidase techniques for identification of neutrophils.

Principle :

This highly metachromatic dye stains neutrophil granules a bright crimson color. In contrast, granules in eosinophils, monocytes, and immature cells of the neutrophilic granulocytic series stain poorly or not at all. Granules in basophils stain deep blue.

Reagents :

1. FAA fixative
2. NEUTROCOLOR stain, in powdered form.
3. Buffer.
5. Cyanoacrylate adhesive.

Procedure :

1. Fix coverslips or slides containing peripheral blood, buffy coat, or bone marrow in the FAA fixative for 5 minutes.
2. Wash in running distilled water for 1 minute.
3. To one of the bottles containing NEUTROCOLOR stain powder, add 14 mL distilled water, agitate gently, then add 14 mL of the buffer and agitate gently.When reconstituted in this way, this stain is stable at room temperature for 24 hours.
4. Apply the stain to the surface of the fixed coverslips of slides and stain for 10 minutes. The presence of undissolved dye particles in the stain solution does not affect the performance of the stain.
5. Pour the Final Rinse Water into a small beaker. Drain off stain by touching the edge of the slide or coverslip to a piece of filter paper.Rinse the dye from the slide or coverslip by holding it with a forceps and vigorously agitating it for 5 seconds in the Final Rinse water.
DO NOT USE DISTILLED WATER. Use only the Final Rinse Water supplied in the kit.
6. Drain dry, and mount with cyanoacrylate adhesive. Do not use a synthetic xylene based mounting medium since the organic solvent dissolves the crimson reaction product obtained with NEUTROCOLOR.

Results :

Neutrophil granules stain bright crimson. Granules in eosinophils as well as granules in immature neutrophilic granulocytic cells such as in myeloblasts, promyelocytes, myelocytes and metamyelocytes stain poorly or not at all. In bands, a few crimson colored granules are identified. In basophils, granules stain blue. The cytoplasm of monocytes stains pale lavender. Nuclei of neutrophils stain dark blue.