Fwd: NADPHd artifact

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From:Susan Travers <travers.3@osu.edu>
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>Date: Fri, 18 Feb 2000 10:23:31 -0500
>To: histonet@pathology.swmed.edu
>From: Susan Travers <travers.3@osu.edu>
>Subject: NADPHd  artifact
>I am writing to ask if anyone has seen and has a solution to an artifact 
>that we have seen in our NADPHd material.  It appears as a small, 
>elongated crystalline precipitate that appears to be on the surface of the 
>tissue. Our technique for doing the NADPHd is the standard one using .1% 
>bNADPH & .01% nitroblue tetrazolium in PB. The only twists are: that the 
>tissue is fixed with PLP , and we double-label the tissue using DAB 
>immunohistochemistry (first NADPHd, then ICC)
>We have tried to trouble shoot by (1) filtering the staining solution-- no 
>help, (2) doing the reaction in the dark (inconvenient and no help), (3) 
>exploring the effects of using ETOH to dehydrate or not-- this is 
>interesting-- with ETOH, which is our standard procedure the artifact 
>looks crystalline, without ETOH-- looks like very small spherical 
>globules, (4) using xylene or HEMO-D to clear-- no effect, and 
>(5)coverslipping with permount versus cytoseal (no effect).
>The only other hints are that we never see the artifact on wet, 
>uncoverslipped sections, and occasionally it seems that the artifact gets 
>worse the first few days after coverslipping; also the artifact does seem 
>worse in the double-labeled tissue.
>Any advice would be greatly appreciated.
>Susan Travers

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