Hi to all developmental biology experts,
I would highly appreciate your advice regarding the shipping of mouse embryos (age E12), from US to Israel. After transportation embryos should be embedded in OCT, cryosectioned and used for Immuno.
What makes this specific situation problematic is that frozen or refrigirated shippment is not an option! Only option is to ship them on room temperature. After the PFA step, what would be the best medium to transport embryos in - PBS, 15,30% Sucrose or something else?
Paraffin embedding is not an option also.
Thank you Lale.
It will take couple of days for transport, that time in PFA would make them unusable for cryosectioning afterwards, even if the epitopes are not ruined. PBS can be better option then sucrose/pbs, thanks.
ya, you don't want to over fix with PFA. A couple of days might be OK, but 4ºC is much better (you could do a week in 30% sucrose at 4ºC after fixation). Try to minimize the time at RT to a couple days max. You will be able to tell by looking at the tissue if it is going bad. good luck!
We've found that 1% PFA is very good for shipping. 3-4 days in 1% PFA for an E12 embryo still gives good results for RNA in situ and IHC. We routinely ship/receive tissue in 1% PFA for cryosectioning. However, they're ideally shipped on cold packs. Perhaps a day shorter at room temperature would be ok, but I don't have a feeling for 1% PFA at room temperature. Otherwise, fix as normal, then ship in PBS. I agree with the previous answers that sucrose is not the way to go.
I myself received and shipped mouse tissues in PBS after fixation. In case of mail delay I think it's the safer. I won't do in PFA to avoid over fixation. I don't like much long wait in sucrose at RT in case of bacterial contamination.
1% PFA for shipping, interesting. Its good to know for the future. Thanks a lot Brian!
Thank you Malki, as we all agreed tissue is already on the way in PBS with 0,01% Sodium azzide, which is know to be a Buffer Storage Solution.
I will let you know how it looks upon arrival.
@ Vukasin: Dear Vukasin, nice to read that you arrived at a practicable solution for the "shipping" ('air mail', I guess). What I would like to know (only from a scientific point of view): you and the commenters wrote only: "1% PFA" - "in PBS" or placing tissue after primary fixation with PFA "for shipping in PBS" ....anyone defined the composition of PBS? (molarity, tonicity, pH?).... I also second the opinion that "shipping at RT" in 1% -buffered- PFA is a good option. Important IMHO certainly is the right molarity/ionic concentration and pH of the buffer solution when shipping at RT is necessary or the only practical solution. Needless to add that shiping with embryos IN (1%,..x.%) PFA solution might pose security questions tto be solved (e.g. double packaging for absolute tightness, "Hazard", "Medical or diagnostic goods"). Best wishes and regrads, WM
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