Description

A set of three different stains for rapid staining of smears for cytological and haematological microscopy. Results will be similar to Romanowsky-stained smears.

Reagents

A. RAPI-DIFF II A Fixative Solution – 500 mL or 250ml (contains thiazine dye in methanol)
B. RAPI-DIFF Solution B – 500ml or 250ml (contains Eosin Y dye in phosphate buffer)
C. RAPI-DIFF Solution C – 500ml or 250ml (contains polychrome methylene blue in phosphate buffer)

Preparation

• Dispense an aliquot of each solution into one of our Collins with a plastic screw cap. Stain Jars (Order Code: 1625) or something similar.
• Prepare a pH 6.8 phosphate buffer solution (order code: 0626) in one wash bottle (Order Code: 1450).

Staining protocol

1. Prepare thin smears on non-greasy slides and air dry.
2. Immerse in Solution A for at least 5 seconds or more.
3. Transfer, without rinsing or drying, to Solution B by immersing and removing slide 5 times allowing 1-2 seconds between dives.
4. Drain excess stain on absorbent paper and rinse with pH phosphate buffer solution 6.8 for a bottle of wash.
5. Transfer slide to Solution C and repeat staining as for B above.
6. Rinse the slide in pH 6.8 phosphate buffer solution and allow it to dry before examination under the microscope.

RAPI-DIFF II Notes

• The number of dips in the spots can be varied to provide different red/blue intensities staining
• When in use, dyes should be stored in suitable covered containers to avoid evaporation (for example Coplin staining bottles with screw caps).
• Washing in pH 6.8 phosphate buffer solution is recommended for best results. Distilled water or tap water can be used in an emergency with some colour loss quality.
• Transfer of Solution B to Solution C will cause deterioration of staining performance over time due to an interaction between solutions that result in the need to increase staining time in solution C. This can be minimized if the slide is washed with phosphate buffer solution pH 6.8 before transfer to solution c
• Solution A may be out of stock before Solutions B and C. We supply each separate solution.