Hi there! I'm performing IHC experiments on PFA-fixed human day 5 embryos (blastocyst stage). After the staining steps, I perform washing in 0.1% Tween-20 in PBS. Although it is usually fine, sometimes embryos tend to attach to the bottom of the dish during washing. The dish we have for washing are not low-adhesion treated (24-well Falcon™ 353047 – Fisher Scientific (10048760)).
I wonder whether I could supplement the washing buffer (0.1% Tween-20 in PBS) with BSA in order to prevent sticking? Any suggestions also about the concentration of BSA to use? Would this change be innocuous for the rest of the protocol or may induce changes that require further optimization? e.g. antibody concentration.
Thank you!
I saw sticky mouse early embryos when I washed them with PBS in 96-well plate (shouldn't be much different to the falcon dishes). I would suggest you check your Tween-20 stock. PBSTw 0.1% is a good condition so I don't think that would be a problem. Maybe borrow some fresh Tween-20 from other labs will solve your issue.
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