I am interested in performing histological analysis (H&E staining, immunostaining etc) on human skin explant ex vivo and wonder which fixative is the best to preserve the overall structure and morphology of the native skin (e.g. intact dermal-epidermal junction, clear differentiated layers of epidermal keratinocytes).
There is a paper (doi: 10.1097/01.pai.0000142159.75540.e0) that compares between different fixatives but they are using reconstructed human skin equivalents, which might be different in terms of their compacity and resistance towards shrinkage during fixation.
Please let me know if any of you had experience dealing with human skin explant fixation and which fixation method yields the best result.
Thanks!
Subhash C. Juneja Thanks for your reply! Do you have any publication regarding ex vivo human skin explant that I can refer to?
I have plenty experience of fixing probably every tissue including skin in general. You can also talk to some pathology lab in your institute or in any institute in USA, give them a call or e-mail, they will answer you. Most of people fix in 10%NBF. 4% PFA is considered slightly better.
As far as i understand, Formalin is thought to preserve tissue morphology better than alcohol fixatives.
Do not use old PFA. Try to make fresh and use within 3/4 days and store in refrigerator for next use. Remember it is carcinogenic, use mask and prepare in a suitable hood. Or you can buy from company called EM sciences (They supply readymade as 8% or 16% in sealed capsules). I used these capsules. Dilute to 4% with PBS. If it is inconvenient for you to arrange PFA, than10% neutral buffered formalin will do the job also. Good luck
I think...store tissue in Bouins fixative for overnight after that give changes with 70% alcohal to remove excess stain. 10%formalin can also b used. These fixative are best for rat tissue and I think will also work for human skin. All the best you can try.
Before placing in fixative tissue important point to be remembered that firstly wash the tissue in normal 0.9% saline and immediately store in bouins fixative in case of vivo study I think. All the best
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