Hey everyone,
I have done a lot of staining for cells on regular 2D surfaces (actin, microtubules, DAPI, etc). But I am now trying to stain cells that are embedded in 3D gels. I looked up papers online/previous posts on here and the process seems to be very similar to regular staining (fixing in 4% PFA, etc). However, I am just a little confused regarding how the aspiration works? Normally, when the cells are in liquid media, I would start by aspirating the media out and then adding the PFA. But in the case of gels, what is it that I am aspirating? Do I basically aspirate whatever liquid component of the gel drains out? And then I guess I have to hope that the PFA (and other subsequent buffers and antibodies, etc) just diffuse through the gel and reach the cells inside? Any help would be very much appreciated!
Thanks,
Appy
Here is a protocol I found online for staining gels, you should be aspirating the cell media from the gels before adding the 4% PFA. This protocol recommends carefully aspirating and leaving about 1/4 of your media in the gel well (25uL in a 96 well plate) so as not to risk damaging the gel.
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