[Histonet] plastic sectioning for light microscopy-HELP!! (H&E staining)

Akemi Allison-Tacha akemiat3377 <@t> yahoo.com
Fri Jan 25 15:42:22 CST 2008

Hi Again,
Sorry, I forgot our method for H&E's.  Generally
speaking, people would use Gills hematoxylin and Eosin
or eosin/phloxine.  Our pathologists didn't like the
look of Gills III.  In 1980, after doing extensive
R&D, a light when on in my brain.  Our nuclear
staining on our GMA PAS's were beautiful.  We oxidized
in 10% periodic acid for 10 minutes for PAS.  So, I
tried doing the same with the H&E's on GMA's.  The
pathologists loved the results and said it looked like
a regular H&E.

I gave several workshops on this subject regarding GMA
H&E's and special stain,s but I can't find my hand-out
material.  Sorry, it was back in the 80's.

Here is the procedure from the cobwebs of my brain:

1. After adhering section to slide, place GMA slide
directly into 10% Periodic Acid for 10 minutes.
2. Rinse in tap water for 1-2 minutes.
3. Place in Harris Hematoxylin for 15 to 20 minutes.
4. Rinse in tap water.
5. Quickly dip in 0.5% acid alcohol.
6. Rinse in tap water.
7. Blue in freshly prepared ammonia water.
8. Rinse in tap water for 10 minutes.
9. Counterstain in Eosin/Phloxine for desired length
of time.  You can start with  1-2 minutes.
10. Dehydrate through graded alcohols quickly.
11. Clear and coverslip.

Hope this helps.

Akemi Allison-Tacha BS, HT (ASCP) HTL
Client Services Manager
PhenoPath Laboratories
551 N 34th St., #100
Seattle, WA 98103
(206) 374-9000
akemi <@t> phenopath.com 

--- Danielle Crippen <dcrippen <@t> buckinstitute.org>

> Dear Histo-experts,
> This is a first for me as I have only done ultrathin
> sectioning for EM.
> I need 0.5-1um serial sections for light microscopy.
>  My main problem at
> this point is transferring the ribbons to a glass
> (plus) slide.  No
> matter how I try to attempt the transfer, the ribbon
> comes apart and/or
> sections are lost or badly wrinkled.  Any and all
> suggestions in this
> area are very welcome!!
> Additionally...once I actually get the sections onto
> the slide (which
> feels like it might be cause for a celebration at
> this point!!)...what
> are the most recommended H&E staining methods (these
> samples are
> embedded in EPON)?
> Again...I am a complete novice at plastic sectioning
> for light
> microscopy...so any suggestions are welcome...even
> the most trivial:-)
> A thousand thanks in advance!!
> danielle
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Akemi Allison-Tacha, BS, HT(ASCP)HTL
Phoenix Lab Consulting & Staffing
Tele: (925)788-0900
E-Mail: akemiat3377 <@t> yahoo.com

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