[Histonet] Staining Glycol Methacrylate

[Jurcisek, Joseph]

Leslie,

	I used to do a lot of JB-4 work.  We had an HEA stain
(Hematoxylin, Eosin, Azure) that we really liked.  Here is the protocol

Reagents
Hematoxylin (we used Harris)
Eosin		0.5% buffered Eosin (buffered in 0.01M phosphate buffer
pH 5)
Azure		0.1% buffered Azure II (buffered in phosphate buffer
pH2.8)

Procedure
Hematoxylin					1.5 min
tap water					rinse
Eosin						20 seconds
di H2O					rinse
di H2O					rinse
Azure						1.5-2 min
di H2O					rinse
di H2O					rinse
Ethylene Glycol monomethyl ether	1 quick dip
di H2O					rinse

	Since JB-4 is miscible in water, there is no need to hydrate
through graded alcohols.  I also have a staining protocol that used
microwaves and create a very vibrant stain.  Unfortunately it is buried
in ancient files right now and will require some digging.  If you are
interested, let me know and I will do it.

Joe

Joseph A. Jurcisek
Senior Research Associate
Center for Microbial Pathogenesis
Columbus Children's Research Institute
700 Children's Dr.
Columbus, OH 43205-2696
(614) 355-3521
fax (614) 722-2818
jurcisej <@t> ccri.net

-----Original Message-----
From: Leslie D Duncan [mailto:leslie.duncan <@t> bms.com] 
Sent: Friday, March 24, 2006 1:28 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Staining Glycol Methacrylate

Hi,

I am experimenting with glycol methacrylate embedded rodent testes.  I 
am looking for a good H&E staining protocol to use that will give 
results comparable to paraffin-embedded tissue (per request of my 
management).  We have dehydrated on the tissue processor to 100% 
ethanol, ilfiltrated and embedded with JB-4, and sectioned at 1-2 
microns.  The sections are placed on Poly L-lysine slides.  Any 
protocols, ideas, and/or suggestions will be greatly appreciated!

Thank you,
Leslie Duncan
Bristol-Myers Squibb, PRI