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Transwell migration assay (Boyden chambers) and DAPI staining


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#1 Tif

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Posted 14 June 2013 - 11:45 AM

Hi all!

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!

#2 Inmost sun

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Posted 18 July 2013 - 03:08 AM

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...




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