There is a web site with great photos of muscle stains, which includes
NADH. However, I don't know of any site showing rat brain with
NADH.

The site is: Neuromuscular Disease Center from the Washington
University School of Medicine, St. Louis, MO.

If you want to see the entire site, go to:

http://www.neuro.wustl.edu/neuromuscular/index.html

If you want to look at the stains, or find staining procedures, go to:

http://www.neuro.wustl.edu/neuromuscular/lab/mbiopsy.htm

There is information on how to do a biopsy, how to do
stains, how to read results, etc.

For the stains, Click on Stains.

Click on NADH (or any other muscle stain). You will get
the procedure for that stain. Towards the end of the procedure,
you can click on Results. This will give you photos of the
stain, with normal and diseased muscles.

I just found this site a couple of weeks ago. I'm planning on
adding it to the Histotech's Home Page, under Peggy's Links.
http://www.histology.to
(Be patient. Steven and I are updating the dead links, and
will be adding new sites. So keep watching our site.)

Peggy A. Wenk, HTL(ASCP)
William Beaumont Hospital
Royal Oak, MI 48073

----- Original Message -----
From: "Anila Syed" <syedab@totalise.co.uk>
To: "Julie Maier" <Julie-Maier@mail.omrf.ouhsc.edu>
Cc: <histonet@pathology.swmed.edu>
Sent: Friday, August 03, 2001 9:46 AM
Subject: Re: NADPH-diaphorase


> Hi, Julie,
>
> Many thanks for your response,
> I have just done a test run on some sections and the results were
horrible.
> They were completely brown!!! I haven't cleared them yet, but I am not
very
> hopeful. I though they were supposed to be blue ?? No-one has any photos
of
> any do they?
>
> Many Thanks,
>
> Anila
> ----- Original Message -----
> From: Julie Maier <Julie-Maier@mail.omrf.ouhsc.edu>
> To: 'Anila Syed ' <syedab@totalise.co.uk>
> Cc: <histonet@pathology.swmed.edu>
> Sent: Friday, August 03, 2001 1:59 PM
> Subject: RE: NADPH-diaphorase
>
>
> > Anila-
> >
> > I did NADPH diaphorase for two years staining for nNOS in rat brain PVN.
> We
> > were using 30um cryosections which were placed (floated) in diaphorase
> > solution immediately after being cut (no cold incubation), incubated 1
> hour
> > at 37C, then rinsed in PBS prior to floating onto slides.  Our PBS was
pH
> > 7.4.  Hope this helps!
> >
> > Julie
> >
> > -----Original Message-----
> > From: Anila Syed
> > To: Histonet
> > Sent: 8/3/01 5:21 AM
> > Subject: NADPH-diaphorase
> >
> > Apologies if this question has been asked before,
> >
> > I am using the NADPH-d stain for the first time, staining free floating
> > 50um
> > sections of brainstem.
> > I have been given a protocol for muscle and am filling in the details
> > from
> > websites and asking around.
> > I hope that a kind individual can answer these questions that i have:
> >
> > Some ppl use Tris buffer and some use PBS and it doesn't say what pH the
> > Tris buffer is at. Can I assume that the actual buffer is not important?
> > But
> > the pH is?
> >
> > Also, in soem protocols, there is a cold incubation in buffer before
> > hand
> > and then a 1.5 hour incubation at 37 deg C in NADPH and nitroblue
> > tetrazolium and in others there isn't. What is the reason for incubating
> > in
> > the cold? My sections are coming straight form the fridge in any case.
> > Would
> > I have to do this incubation? Is it just to bring it to the right pH?
> > Wouldn't the temperature affect the pH any