Re: staining JB-4 sections

From:Sarah Jones

Hi Michaela,
   I just did some H&E's on GMA sections this week, here is the
procedure I used:

H&E Stain for Glycol Methacrylate Sections
1.	Gill's Hematoxylin for 30-35 minutes			
2.	Rinse in distilled water, three changes			
3.	Blue in Scott's Tap Water Substitute, 3
4.	Rinse in distilled water, two changes			
5.	Stain in Eosin-Phloxine for 6-8 minutes			
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 Substitute			
1 liter distilled water			
10 gm magnesium sulfate, MgSO4	
2 gm sodium bicarbonate			
Mix until dissolved, and filter before use

(0.5% Eosin Y, 0.1% Phloxine in 0.2% acetic acid)
Add 0.6 ml glacial acetic acid to 500 ml distilled water
Add 2.5 gm Eosin Y and 0.5 gm Phloxine

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. 

Ref: Barbara F. Matlaga
Ethicon Research Foundation
Somerville, NJ  08876
The Bulletin SPEP, Vol. IV, No 2
June 1976

If someone could tell me what "The Bulletin SPEP" was or stands for, I
would be interested to know.  

Sarah Jones HT(ASCP)
Dept. of Vet. Anatomy & Public Health
Histology Lab
Texas A&M University
College Station, TX 77843-4458
phone: 979-845-3177
fax:  979-458-3499

>>> Michaela Fr#253#zkov#225#  01/30/03 09:01AM
Dear Colleagues,
I need an advice about staining sections embedded in JB-4 Plus. I embed
this medium and now I need stain my sections. I wanted to use regular
haematoxylin-eosin staining, but it doesn#180#t work. Do you know
about that? Can you recommend me some other staining?

Thank you, very much.
Michaela Fryzkova

Department of Parasitology
Faculty of Science
Charles University
Czech Republic

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