Fwd: NADPHd artifact
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From: | Susan Travers <travers.3@osu.edu> |
To: | histonet@Pathology.swmed.edu |
Reply-To: | |
Content-Type: | text/plain; charset="us-ascii"; format=flowed |
>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.
>
>
>
>'Regards,
>Susan Travers
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