Re: Re: [Histonet] Good Perfusion...

[TF]

I think many, many labs perfuse animals under deep anesthesia. Notice the word "DEEP". It is hard to judge whether the animal can recover. But we know not because it is an overdose injection.

Then, when you write papers up, you never use "Deep anesthesia", you use "sacrifice". That is some back rules in writing papers, but it make some European reviewers better.

To have good fixation on inschemic area, post-fixation at room temperature with changes of fixative every 24-48 hours is required.


2009-07-01 



TF 



发件人： Chana de Wolf 
发送时间： 2009-07-01  03:42:30 
收件人： jfish 
抄送： histonet; JR R 
主题： Re: [Histonet] Good Perfusion... 
 
Histonetters,
Perfusion under deep anesthesia is most certainly not unethical NOR illegal,
and, in fact (as mentioned by Jo Dee), it is necessary for optimal perfusion
and fixation -- the intracellular ischemic cascade begins immediately upon
circulatory arrest, setting off a chain of events highly detrimental to
subsequent perfusion. Indeed, ischemia quickly leads to the "no-reflow"
phenomenon, effectively guaranteeing that you are not perfusing all tissues
adequately at all! It is therefore *most* beneficial to begin perfusion with
a beating heart, and every perfusion protocol I have ever worked under
requires it (especially if the tissues are to be used for EM).
I perform around 5-10 perfusions per week *specifically* to study the
effects of ischemia on reperfusion and neural ultrastructure.
Of course, you should check with your institution's particular rules and
regulations, but perfusion begun under anesthesia is scientifically
justified for the reasons mentioned above and should therefore be relatively
easy to have approved by your IACUC.
Sincerely,
Chana de Wolf
On Tue, Jun 30, 2009 at 11:37 AM, Jo Dee Fish <jfish <@t> gladstone.ucsf.edu>wrote:
> Dear Jerry and histonetters,
>
> I don't believe this is unethical or illegal.  It is written into our IACUC
> approved protocols as follows:
>
> Chemical Method of Euthanasia:  "Perfusion under general anesthesia,
> Avertin
> or Halothane induced.  Bilateral thoracotomy."
>
> Could it only be forbidden by your facility?
>
> I wonder if other facilities have rules against such things.  But we insist
> that our investigators use perfusion under anesthsia because the deeper
> tissues are much better fixed and blood removal is more thorough and
> complete by this method.
>
> Jo Dee
>
>
> ~~Jo Dee Fish~~
> Senior Research Technologist
> The J. David Gladstone Institutes
> Co-manager Histology and Microscopy Core
>
> Telephone: (415) 734-2567
> Fax: (415) 355-0824
> E-mail: jfish <@t> gladstone.ucsf.edu
>
> Mailing address:
> The J. David Gladstone Institutes
> 1650 Owens Street
> San Francisco, CA 94158
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of JR R
> Sent: Tuesday, June 30, 2009 10:49 AM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Good Perfusion...
>
>
> No, I wouldn't nick the atrium.
>
> Instead, cut the femoral artery at the groin.
>
> And of course, you cannot perfuse a living, anesthetized animal.  That
> would
> be unethical and also illegal.
>
> Jerry Ricks
> Research Scientist
> University of Washington
> Department of Pathology
>
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