[Histonet] slides for frozens

[Sherwood, Margaret]

To all:

We have been using the same slides for paraffins and frozens.  We have no
trouble with paraffin-embedded tissue adhering to the slides through the whole
staining procedure.  But our frozens (even 5um sections)have been falling off
during manual staining.  The slides are (+) charged.  We used to use slides from
Fisher, but they are so expensive, we changed to an independent vendor (half the
price).

Our procedure for frozen H&Es has recently changed:
1. Fix in 95% ETOH       1 min.
2. Hematoxylin           5 min.
3. Acetic acid H2O	 2 quick dips
4. Wash-running H2O      10 seconds (10 dips)
5. Bluing reagent	       10 dips
6. Wash-running H2O      10 seconds (or 10 dips)
7. Eosin	             5 seconds
8. Dehydrate-95% ETOH    10 dips each
9. Dehydrate-100%ETOH    10 dips each
10.CitriSolv             10 dips each
11.Coverslip

The tissue seems to fall off in the bluing reagent (we order from Mossberg).

Can anyone help us?  Suggest anything--what slides you are using?  We have had
the same problem with special stains (i.e. Oil Red O for fat on frozens). 

Thanks!
Peggy

P.S.  I use the same slides for my 1um plastic embedding which I dip in water
rinses and have no problem with the tissue falling off.


Peggy Sherwood
Lab Associate, Photopathology
Wellman Center for Photomedicine (EDR 214)
Massachusetts General Hospital
50 Blossom Street
Boston, MA 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood <@t> partners.org



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