I'm using 10 and 25% sucrose buffer for cryopreservation after fixating the samples in PFA. I wonder how pure the sucrose should be? Does it need to be 99% pure or so?
Hello Sahar,
As far as my knowledge concerned there is nothing wrong in using low purity sucrose for cryo-preservation, but the thing is if u go for high purity/ ultra-pure sucrose the results will be much much better and u can minimize the source of contamination if your using it for research purpose. It always better to go for high purity chemicals for any procedure that involves biological material.
Regards,
Sanketh D.S
Thanks so much Sanketh. That was indeed a good point; to go for high purity chemicals for biological materials.
Dear Sahar
As Sanketh said, don't worry about the purity of sucrose, unless the condition of its stocking was not good, or may be mixed with something else during laboratory activities(handling, storage,spatuls,balances,....).
According to my experiences on fish sperm cryopreservation, your not satisfied results might be related to the fixing material and methods, because when you fixed the cells they are already affected with a new and not normal environment.
Good luck
Shahrouz
Thanks Shahrouz for your thorough explanation. One more question, do you know if the epitopes can be affected by PFA or even sucrose? I got no signal for a particular antibody that I used (and IO know the antibody should work), now I'm wondering if it could be due to the steps before sectioning, i.e. PFA or sucrose
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