It could be doable with some effort but I feel that it would negate the whole purpose of matrigel culture/assay, which is meant to provide three-dimensional matrix environment for cells to migrate with changes in shape and physical appearance/membrane properties. These changes could be best captured by microscopy and when dissociated of the matrigel majority of these changes will be lost. 

After being exposed to warm/room temperatures, matrigel will not melt completely upon cooling,. I do admit that I did try to harvest the matrigel exposed cells, which are not the same cells anymore, but definitely useful for WB and RT-PCR, and worked fairly well. If you are really planning FACS for characterizing some proteins, I wouldn't discourage you but WB is a better choice though!

Thanks for your answer Yagna :) The reason for FACSing them is to assess the number of migrated cells in a more unbiased manner than selecting "random" areas to count by microscopy. But indeed, if I risk clogging our FACS, I'll go for the in-gel counting instead.

Cheers,

Annette

Hey Annette,

dou you only want to count cells in the FACS? If so, I think the necessary preparation to generate samples suited for flow cytometry isn't worth it and I doubt it will work on the fly. If you want to do immunostaining I've heard that fixation and slice preparation of matrigel is possible and that certain antibodies can reliably diffuse into the structures. However that sounds like an awful lot of messy work so I've never tried that.

 I agree with Yaschar, it would be quite messy and may not be unbiased anymore.

Just for counting, I would suggest loading cells with a cytoplasmic fluorescent dye, and following migration you can easily/reliably count them compared to without any staining. This does not fixing n sectioning, would enable imaging the cells migrating in three dimensional directions by confocal microscopy, and you would be able to see the volume of space covered by migrating cells (that could be constructed by an appropriate software, if you really want to go that far, really worth it!).

Another advancement that you could consider is to express GFP or any FP of your choice and do the imaging, so that you don't have to deal with choosing an appropriate dye, which itself needs some mind-storming.

Hi Annette, I think the IncuCyte machine of Essen Bioscience can help you. We just bought one. Best regards, Evelien