Re: [Histonet] Murine perfusion for whole autoradiography

From:Geoff McAuliffe

Hi Jamie:

First off is there so much I125 in the blood that it will obscure your 
results? Is someone going to complain that washing the blood out skews 
the "true physiology"?

Secondly, if perfusion pressure is too high it will damage the blood 
vessels but in that case I think any subsequent procesing, washing, etc. 
would remove the blood and any bound I125? You would have to make a 
pretty big hole to push antibody molecules through.

How high is too high? I suspect blood pressure in a mouse is not much 
different from any other mammal, ie humans, and such information is 
available. Converting inches of water (how high to hang the perfusion 
bottle) to millemeter of mercury is just arithmetic. 1 inch = 25.4 mm 
and mercury has a specific gravity of 13.6. You could also buy a 
peristatic pump and hook up a pressure gauge to the output side 


Jamie E Erickson wrote:

>HI All,
>               I saw this posting of perfusion for mouse brain and had to 
>ask a similar question. 
>I am working on doing biodistrubtion of whole mice with I125 labeled 
>antibodies. The question I have is I want to perfuse out all the 
>radioactive blood from my mice before sectioning the whole mouse  so that 
>I can look to see where the binding might be and have little radioactivity 
>due to blood content.  If the pressure in the perfusion is to high this 
>may push my radioactive antibody by pressure my  into an area that is not 
>where it would have gone in-vivo without perfusion giving me a false 
>positive location.
>      So is there a way to mimic physiological pressure in a mouse so that 
>this would not be an issue. How high would you hang the perfusion bag for 
>a mouse???
>Any thoughts???
>Jamie Erickson
>Sr. Research Associate 
>Department: DSMP
>Abbott Bioresearch Center
>100 Research Drive
>Worcester, MA 01605-4341
>FAX: 508-793-4895
>Histonet mailing list

Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029

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