Planning on doing transcardial perfusions to clear/fix brain tissue in postnatal day 0-1 rat pups. I have never perfused anything that young/small, so I am looking for tips. Are there any special tricks, other than reducing size of needle, volume of solutions, and going slow? Advice would be appreciated.
hi pamela
may be also using body warm perfusion solution would help.. or you may even try to fix the brain without perfusion as a parallel experiment to check if the perfusion is necessary for your case..i wish you good luck
Dear Pamela,
This work is very simple. After anesthesia with an appropriate method you should place the rats pup on ice. For brain histological studies fixative solution containing 4% paraformaldehyde in 0.1 M phosphate buffer (pH 7.4) is acceptable. You can use this solution (cold) in 10 mL syringe and 26-28 needles. For pups at postnatal day 1, without opening their chest you can insert the needle into their heart and inject at least 5 mL fixative solution. If the brain vessels were clear and the brain tissue was consistence your perfusions have been corrected. For older pups you should open their chest and perfusion them according to the standard protocols. (for Example you can see the attached protocol)
Good Luck
http://www.jove.com/video/3564/whole-animal-perfusion-fixation-for-rodents
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