RE: [Histonet] SDH protocol


I guess the attachment didn't come through..........Here is the procedure:



Prepare this substrate solution at least on e hour before incubating and preheat in 37C.

0.1M Sodium succinate			15ml
0.1M Phosphate buffer pH 7.6		15ml
Nitro Blue Tetrazolium (refrigerated)	15mg


1.	Frozen sections are cut 6 – 8 microns and stay in the cryostat until ready to    
2.	Place slides in coplin jar and incubate for 30 minutes at 37C. 
3.	Rinse slides with distilled water and coverslip from water with Aqua mount.

Ref. Histochemistry, A.G. Parse, page 910 


Beatrice DeBrosse-Serra
Allergan, Inc.
2525 Dupont Drive RD-2A
Irvine, CA 92612

-----Original Message-----
From: [] On Behalf Of Lewis, Sarah
Sent: Friday, December 09, 2005 7:19 AM
To: HISTONET (E-mail)
Subject: [Histonet] SDH protocol

Fellow Neuromuscular Labs!!
I am in need of a Great SDH protocol.  I have been struggling with this stain for a long time.  I purchase new NBT and it works pretty well for a few weeks.  Then is starts fading into nothing within 1month.   I use a protocol of on the WashU website (St. Louis MO). It works better than any other I have tried... but still very pale, However my pathologist said that the SDH's in the past have always been more blue.  Mine come out purple/ pink.  I have also used the protocol from past histologist in this lab with no luck.  I must be missing some small technique that isn't written in the protocol.  Please help!!!       

Sarah E. Lewis 
Childrens Research 
Center for Gene Therapy 
700 Childrens Dr Rm WA3112 
Columbus OH 43205 

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