Deep Blue Stain

These images represent a new stain based on picro aniline, Ziehl’s fuchsin, and acetic acid. The stain is useful in dermatopathology because of its utility in differentiating squamous epithelium keratinization (yellow/brown) and lymphocytes (dark red). It is useful in gastric biopsies to differentiate metaplastic glands.

Protocol

Solution A: 100 mL distilled water + 1 drop Ziehl’s fuchsin (from Ziehl-Neelsen stain) + 1 drop 9% acetic acid

Solution B: 100 mL distilled water + 1 drop 10% formaldehyde + 1 drop 9% acetic acid

Solution C: Picro aniline

1. Dewax and rehydrate.
2. Wash 5–10 seconds in distilled water.
3. Put in Solution A for 2–5 minutes.
4. Wash 5–10 seconds in distilled water.
5. Put in Solution B for 5 minutes.
6. Wash 5–10 seconds in distilled water.
7. Put picro aniline on sections for 5 minutes.
8. Wash slides until blue color appears.
9. Put in xylene for 3 minutes.
10. Mount coverslips.

Submitted by Dmitry A. Zinovkin, pathologist at Gomel State Medical University, Gomel, Belarus.