Re: RE: Re: [Histonet] Fixation of the damaged brain tissue

[TF]

Thanks, Dont you think high pressure during perfusion leads to tissue swelling and disrupted morphology?


2009-01-13 



TF 



发件人： Charles Scouten 
发送时间： 2009-01-13  00:02:40 
收件人： tifei <@t> foxmail.com 
抄送： histonet 
主题： RE: Re: [Histonet] Fixation of the damaged brain tissue 
 
Formaldehyde penetrates very slowly, that is why we do perfusion in the first place.  To get fixative everywhere very quickly, since autolysis and breakdown starts almost immediately with anoxia.  The tissue around the wound is decaying quickly, and not getting fixed fast enough. 
Consider using a higher pressure on the perfusion to force washout of the extra vascular space.
Cordially,
Charles W. Scouten, Ph.D
Innovation Manager
Biosystems Division
Leica Biosystems St. Louis LLC
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United States of America
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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of TF
Sent: Monday, January 12, 2009 9:42 AM
To: Geoff McAuliffe
Cc: histonet
Subject: Re: Re: [Histonet] Fixation of the damaged brain tissue
4% PFA
so u mean I should expose more of the damaged site (it is at the bottom of the brain, or olfactory bulb) and immerse the whole brain in a larger volume of fixatives?
I think PFA penetrate quite fast, and I took all the skins & muscles off the skull before put the who brain with skull into PFA for 1-2 days.
It should work...
The perfusion is not well in many conditions - in my injury model, part of the blood supply was interrupted.
2009-01-12 
TF 
发件人： Geoff McAuliffe
发送时间： 2009-01-12  22:59:22
收件人： tifei
抄送： histonet
主题： Re: [Histonet] Fixation of the damaged brain tissue 

Hi TF:
Your problem should not be happening.
Sounds like poor perfusion followed by immersing the whole brain (with the wound deep in the interior) in a small volume of fix at 4 degrees C.
What fixative are you using?
Geoff
TF wrote:
> Hi, I just want to fix the damaged brain tissue (2-48 hours after I made the injury).
> Because the injury damaged the surrounding blood supply, and after perfusion I normally got a mass of soft, shapeless tissue surrounding the injury site...
> I have also tried post-fixation of 24-48 hours before removing the brain from in situ. But it does not help a lot.
> Can anyone provide some suggestions?
>
> 2009-01-12
>
>
>
> TF
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Neuroscience and Cell Biology
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