Transwell migration assay (Boyden chambers) and DAPI staining - (Jun/14/2013 )
Does someone have any experience with the boyden chambers?
I performed this assay and, in the last step, I stained the migrated cells with DAPI after fixing and permeabilizing them. I looked at them with the fluorescence and I could clearly see the blue dots of the nuclei. The problem is that I could not see cells with the bright field, so I am afraid that what I observed could be the aspecific of the membrane pores. Could this be possible? Should DAPI stain only the DNA ? Could the blue dots be migrating cells that are still stacked into the membrane pores?
Thank you very much for your help!
do you know if your cells are attaching to the plastic cell culture chambers? if so, migrated cells should be detectable in bright field microscopy; to increase migration you may coat the bottom of wells with extracellular matrix proteins...
the DAPI stain is just to ease counting of cells...