Normally tissue is fixed in 4% PFA (or animal perfused with 4% PFA before taking tissue from animal), than the tissue passed thru 15% sucrose and than in 30% sucrose before freezing in OCT blocks. Why sucrose concentration is chosen at 30%?
We learn that sucrose will decrease the cell deformation. I think it will also limit the swelling of your tissue during the fixation processes. An old article (Bahr 1957) told that the deformation was inversely proportional to the sucrose concentration.
For best cryoprotection of fixed tissue, you need to do sucrose infiltration.
Dear Subash,
There were other facts too (why 30% sucrose)....
Proposal:
Search ResearchGate repository (for Authors, Publications, Questions etc.etc. as appropriate by using its "SEARCH" function (full text search possible), insert your search word, search phrase or text into the rectangel , click the loupe and wait a second....then choose from auithors, publixcations, questions....
eg.: ' cryoprotect ' results in creating an URL= https://researchgate.net/search.Search.html?type=question&query=cryoprotect
Hi Subhash, as Shambhu mentioned, it helps to prevent freeze damage through (water-) ice crystal formation. Sucrose solutions of 1.8M or higher freeze glass like (vitrification, i.e. no formation of damaging ice crystals) no matter how slow the freezing takes place (Griffiths, McDowall et al. 1984; McDowall, Chang et al. 1983).
Hi Zaara, I believe you want to make 30% sucrose, right
To 12 gm sugar add PBS till total volume is 40 ml
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