[Histonet] GRAM Stain Timing Question

[Maxim Peshkov]

Sara,
Our protocol for GRAM:
1. Dewax.
2. DW, rinse (several secs).
3. 1% Aqv. Gentianviolet(+Tween20 5 drops in
100mL), 1 min.
4. DW, rinse (several secs).
5. Grams's iodine 1 min.
6. DW, rinse (several secs).
7. Blot section dry with filter paper.
8. ACETONE, 2-4 DIPS, until excess blue dye comes
off of the sections.
9. Tap water several secs.
10. DW with 2-3 drops glacial acetic acid on 100mL,
30-60 secs.
11. 1% Aqv. Neutral red(+Tween20 5 drops;
+2 drops glacial acetic acid in 100mL), 1 min.
12. Blot section dry with filter paper.
13. Acetone, 3 changes, each 10 dips.
14. Xylene, 2 changes, each 10 dips.
15. DPX, coverslip.

All reagents we prepare by itself.
This protocol we doing manually, daily, many
years. It is work great. Pathologists are lucky,
and techs also.

Good luck,
Maxim Peshkov
Taganrog
Russia,
Maxim_71 <@t> mail.ru