Pam,

Without knowing what your staining procedure is, it seems as if you may be
getting uneven staining due to improper deparaffinization and incomplete
hydration.  It does not sound to me like you are having a drying problem.

You may want to use the basic histology procedure for H&E staining you will
find in most textbooks.  These procedures are always a good base to start
developing you own with and it is always good to go back to basics when
there is a need for troubleshooting problems.

Here are some things you may want to try.

1.  Make sure your slides are in 3 changes of xylene for 3 minutes each,  2
changes of 100% for 2 minutes each,  2 changes of 95% for 2 minutes each
and one change of 80% alcohol for 2 minutes, rinse with distilled water
until ripples no longer appear on slides before they go into hematoxylin.
2.  When placing your slides in the dishes vessels agitate them up and down
to make sure the spaces between them are filled.
3.  Most important, make sure your stains are filtered and your
solutions/solvents are clean.

If you do not  have any references,  I would be happy to send you a
procedure from one of my reference books.  E-mail me and let me know.  Or
call me at 800-222-0342x443.

Rande Kline
Technical Services
EM Science






Pam Plumlee <PPlumlee@thermolase.com> on 11/12/98 12:50:30 PM

To:   "'histonet@pathology.swmed.edu'" <histonet@pathology.swmed.edu>
cc:
Subject:  Manual H&E staining problem




Histonetters: I do manual H&E staining on human and animal tissue.
Lately some sections or parts of sections are not taking up the Eosin.
This problem is very intermittent, but very noticeable on slide review.
Sometimes the first level on a slide will stain great and on the third
level the whole right half of the section will not stain.  I've tried
drying the slides longer, alternating the side holder from side to side,
to make sure the paraffin melts correctly.  Any ideas?  Thanks in
advance, pplumlee@thermolase.com