Re: Re: [Histonet] feulgen stain on brain sections

[TF]

Thanks very much.
I will extract the lipids first.


2009-02-10 



TF 



发件人： Geoff McAuliffe 
发送时间： 2009-02-10  00:17:29 
收件人： tifei 
抄送： histonet <@t> lists.utsouthwestern.edu 
主题： Re: [Histonet] feulgen stain on brain sections 
 
Dear TF:
Schiff reagent reacts with the lipids (the aldehydes in the lipids) in 
the frozen sections so the whole section is pink. You could try 
extracting the "offending" lipids with alcohols and xylene, then 
rehydrate to water and run the reaction again. A better plan is to 
search the literature first, the old literature in books in the library. 
Look for Histochemistry by A.G.E. Pearse.
Also 1.5 hours in %N HCl seems excessive, again check the literature for 
the correct time on unfixed frozen sections.
After you solve the Schiff problem the light green in distilled water 
should work fine.
Geoff
TF wrote:
> Hi all, i amusing Feulgen staining to visualize the apoptotic cells on 20 um brain frozen sections.
> The procedure is 5N HCl room temperature for 1.5 hour, wash in cool 1N HCl, wash in water, Schiff staining room temperature 1 h, wash throughly in distilled water, counterstain with 1% light green.
>
> I have two questions here:
> (1) after schiff staining, i can not visualize individual nucleui clearly, but all the section seems to be stained red.
> (2) I use distilled water to prepare 1% light green, am I right? I can not counterstain the section well - or, possibly because the sections are still in red color.
>
>
> 2009-02-07 
>
>
>
> TF 
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>   
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
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