Re: [Histonet] H&E to PAS restaining protocol
Russ Kerschmann MD wrote:
> Can anyone provide a protocol for reprocessing an H&E
> slide into a PAS slide? Can the reverse also be
Easy! Here's a procedure. I'm not happy about
calling this kind of thing a "protocol" because
it needs observation and judgement if it's to
work properly. You should do this job yourself;
do not entrust it to someone else.
1. Remove the coverslip and mountant and
take the section to 70% alcohol. Don't
rush any step of this procedure, and be
especially certain that you have
dissolved out all the mounting medium.
2. Immerse in acid-alcohol until all the
haemalum nuclear (and any other blue)
staining has gone.
3. Rinse in 70% alcohol, then in water.
4. Put in slightly alkaline water until
the eosin colour (pink) has all gone.
(Note. You will probably not be able
to extract eosin from a few strongly
basic materials present in some animal
tissues: eosinophils, Paneth cells and
5. Wash in tap water, then in distilled
water before doing the PAS method.
>How can the risk of losing the tissue be
Keep exposure to alkalinity (Step 4 above)
to a minimum. Hard tap water may be
sufficient, or you can add a small drop of
ammonium hydroxide, or of saturated aqueous
lithium carbonate. High pH removes sections
> Many thanks, and Happy Holidays.
Which holiday(s)? The next one on my calendar is
Nobel Day (Sweden) on 10th December, followed by
Our Lady of Guadalupe (Mexico & Central America)
2 days later. Chanukah isn't until the 19th.
Or do you mean happy Christmas? It's still
25 days away!
Hope this helps.
John A. Kiernan MB, ChB
Department of Anatomy and Cell Biology
The University of Western Ontario
London, Canada N6A 5C1
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