[Histonet] fixation via perfusion

[Geoff McAuliffe]

Hi Wisam:

    I don't have the exact answer you are looking for but I can provide 
some direction. This information was probably worked out in the late 
1950's and/or early 1960's as perfusion fixation emerged as the way to 
fix for electron microscopy. I would suggest looking in Hayat's 
"Principles and Techniques for Electron Microscopy" or Lillie and 
Fullmer's "Histological Technique and Practical Histochemistry" as 
places to look for the original references.
    In my experience, most people use too small a canula and/or too low 
a flow rate for good fixation. The rate of flow you use should 
approximate the cardiac output of the animal in question. As far as 
volume goes, I usually use a 3 times the weight of the animal if I am 
perfusing the whole body, less if you clamp off the lower body and 
perfuse just the head.
    Good luck!

Geoff

Wisam Barkho wrote:

>Hello,
>
> 
>
>We are doing stereotaxic surgeries on rats in my lab. After 2 weeks, we
>fixate the brain via perfusion using 4% PFA (after vascular rinse) and then
>immerse the brains in 4% PFA overnight. My question is where can I find
>information on pump flow rate and PFA volume to use during fixation? Thanks.
>
> 
>
>Wisam Barkho
>
>daufoi <@t> msn.com
>
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
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mcauliff <@t> umdnj.edu
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