Fw: More on [Histonet] Looking for detailed protocol to stain for PAS in liver sections (paraffin)

[Gayle Callis]

Dr. Nasonkin was staining for glycogen (per my question) and via private 
emailing, it was suggested he not use aqueous formalin or PFA fixation,  but 
an alcoholic fixative to retain the glycogen in the liver, do diastase 
digestion to prove it is glyogen, and also process tissues -  starting in 
higher (100%) alcohols to help retain glycogen.   Suggested fixatives were 
Carnoy, Gendre and alcoholic formalin.

If you have any other suggestions for him, please add to the list of to do's 
.

Gayle M. Callis
HTL/HT/MT(ASCP)
Bozeman MT

> ----- Original Message -----
> From: "Igor Nasonkin" <nasonkini <@t> mail.nih.gov>
> To: "Gayle Callis" <gayle.callis <@t> bresnan.net>
> Cc: <mpotok <@t> umich.edu>
> Sent: Tuesday, May 06, 2008 6:35 PM
> Subject: Re: [Histonet] Looking for detailed protocol to stain for PAS in
> liver sections (paraffin)
>
>
> Gayle,
> Yes, glycogen.
> Does it mean that if we fixed livers in paraformaldehyde prepared on PBS 
> we
> lost glycogen? What if we fixed the whole liver, not section on a slide? 
> One
> cannot remove glycogen from the whole liver this way. Thank you for this
> info,
> Igor
>
> Dr. Igor O. Nasonkin
> Research Fellow
> National Institutes of Health/NEI
> 9000 Rockville Pike, MSC 1864
> Bldg 10, Room 10B11
> Bethesda, MD 20892
> Tel: 301-443-7398 ­work
>       617-388-4104 ­cell
> Fax 301-480-1769
> email: nasonkini <@t> mail.nih.gov
> http://www.nei.nih.gov/intramural/nnrl.asp
>
> On 5/6/08 8:08 PM, "Gayle Callis" <gayle.callis <@t> bresnan.net> wrote:
>
>> You did not say what you are trying to see in the liver?  Glycogen? Some
>> other PAS positive tissue component?   If so,aqueous formalin fixation
>> will remove the glyocgen.  An alcoholic fixative helps retain glycogen.
>>
>> Gayle M. Callis
>> HTL/HT/MT(ASCP)
>> Bozeman MT 59715
>>
>> ----- Original Message -----
>> From: "Igor Nasonkin" <nasonkini <@t> mail.nih.gov>
>> To: <Histonet <@t> lists.utsouthwestern.edu>
>> Sent: Tuesday, May 06, 2008 5:24 PM
>> Subject: [Histonet] Looking for detailed protocol to stain for PAS in
>> liver
>> sections (paraffin)
>>
>>> I am looking for a detailed protocol for PAS staining in liver sections
>>> (paraffin-embedded). The 1st set of sections did not work, and we had no
>>> +control since we have not done it. These are 4-6wk mouse liver sections
>>> embedded in paraffin; our lab is experienced in IHC on paraffin sections
>>> so
>>> these were done right. But PAS staining did not work. What could be main
>>> reasons? Any positive control we could use? Thank you in advance,
>>> Igor