[Histonet] RE: H. D., Vol 78, Issue 1, Message: 7

[Keith]

Here's a copy of a procedure that provided consistently adequate results
from my old one-man lab histo days...

For your case, perhaps the slides in question were fixed differently or
improperly(?) I would pull the questionable slides, delicately rehydrate
them, run a control slide, and reprocess them with the following procedure.
In this procedure, the staining agents, dehydrants, and clearing agents and
can be modified to your facility and reagent list.

Keith M. Mikoff, HTL/HT (ASCP)

                              PAPANICOLAOU STAIN (PAP)

          PURPOSE: The following staining procedure is to be performed only
          on spray fixed slides.  It is a routine stain used for any cytol-
          ogy (i.e., PAP smears, FNA procedures, or specimens of a fluid
          nature where cytology is required).

          PROCEDURE:

          1.   Dehydrate slides in 2 changes of isopropyl alcohol, 5
          minutes each.

          2.   Rinse in tap water for two minutes.

          3.   Stain in Harris' Hematoxylin for 2 minutes.

          4.   Rinse in fresh tap water for 3 minutes.

          5.   Place in ammonia water for 1 minute.

          6.   Rinse in fresh tap water for 1 minute.

          7.   Dehydrate in 3 changes of Isopropyl alcohol, 2 minutes each.

          8.   Stain in O-G 6 for one minute.

          9.   Dehydrate in Isopropyl alcohol for 2 changes, 2 minutes
          each.

          10.  Stain in EA-50 for 4 minutes.

          11.  Place in Isopropyl alcohol, 3 changes, at 1 1/2 minutes each

          12.  To clear the slides, place in 3 changes of Histoclear, for 2
          minutes each, and coverslip.