[Histonet] Fwd: MIXED fiber type identification by Immunohistochemistry

[Elizabeth Chlipala]

I think you normally use ATPase at different pH's, which is an enzyme stain, to differentiate muscle fibers.

At pH 10.2 it would stain Type I fibers light and then Type II Fibers dark  (all types IIa, IIb and IIc) the other pH's stratify out the type II fibers.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of arjun ruhella
Sent: Monday, October 22, 2012 11:51 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Fwd: MIXED fiber type identification by Immunohistochemistry

Dear all,

Its been a while i asked for a doubt to all histology experts around US, but i have not been able to get much reply (only 1 :(). I thought my doubt/e mail might have been missed by some, so sending it again.

I would really appreciate if anybody could guide me in my problem.

Hope to hear people's advices this time.
Thanks

Best,
Arjun


Begin forwarded message:

> From: arjun ruhella <ruhella.arjun <@t> gmail.com>
> Date: October 2, 2012 8:34:59 AM EDT
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: MIXED fiber type identification by Immunohistochemistry
>
> Greetings to all,
>
> In immunohistochemistry, I am encountering confusion in designating muscle fibers (based on staining color intensity) to a particular fiber type.
>
> This problem is particularly bothersome when i encounter the fibers stained with 2 different colors, one color has good intensity while other seems to be of light intensity. I don't have any criterion whether to consider this light staining as actual staining, which would classify the fibers as MIXED fibers or should i consider this light staining as background/non specific staining in which case the fibers will be classified as PURE fibers.
>
> Does anybody can suggest any criterion/tips they have been using to identify muscle fiber type as correctly as possible? I can understand that identifying MIXED fibers is subjective in immunohistochemistry, but it would be of immense help if i could get to know of any objective suggestions.
>
>
> Animal= spinal cord injured Rat
> Muscle = Soleus
> Section Thickness = 10µm
>
> I will really appreciate the help!
>
> Thanks, hope to hear from ppl soon!
>
> BEST,
>
> AR

Best,
Arjun

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