Re: [Histonet] H&E staining on GMA plastic

From:"Sarah Jones"

Hi Geoff,
  Here is the one I use.    Sarah

H&E Stain for Glycol Methacrylate Sections

1.	Gill's Hematoxylin for 30-35 minutes			Ref:
Barbara F. Matlaga
2.	Rinse in distilled water, three changes			Ethicon
Research Foundation
3.	Blue in Scott's Tap Water Substitute, 3
minutes		Somerville, NJ  08876
4.	Rinse in distilled water, two changes			The
Bulletin SPEP, Vol. IV, No 2
5.	Stain in Eosin-Phloxine for 6-8 minutes			June
6.	Rinse in two changes of 95% alcohol
7.	Rinse in two changes of Abs. alcohol, allow sections to remain
in the last Abs. for one minute
8.	Clear and mount sections in a synthetic mounting medium

Scott's Tap Water Sub.			Eosin-Phloxine
1 liter distilled water			(0.5% Eosin Y, 0.1%
Phloxine in 0.2% acetic acid)
10 gm magnesium sulfate, MgSO4	Add 0.6 ml glacial acetic acid to 500 ml
distilled water
2 gm sodium bicarbonate			Add 2.5 gm Eosin Y and
0.5 gm Phloxine
Mix until dissolved, and filter before use

Hematoxylin (half-oxidized from Gill)
Add ingredients in order:	365 ml distilled water, 125 ml ethylene
2 gm anhydrous powdered hematoxylin, 0.2 gm sodium iodate (Na2IO3),
weigh out accurately.
40.25 gm aluminum sulfate, Al2(SO4)3*18H2O, 0.5 gm anhydrous citric
acid.  The stain can be used
after 1 hour of mixing.  Filter through Whatman #1 filter paper before
using for the first time.  

Sarah Jones, HT(ASCP)
Histology Lab
Dept. of Veterinary Integrative Biosciences 
College of Veterinary Medicine
Texas A&M University
College Station, TX 77843-4458
phone: 979-845-3177
fax:  979-458-3499

>>> Geoff McAuliffe  01/06/05 6:00 PM >>>
Dear List

    A colleague wants to do some H and E staining on zebrafish embryos

embedded in JB4+ plastic. Does anyone have a good protocol they can 
share? Thanks in advance!


Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029 

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