Cells Detach After Transfection - (Sep/19/2013 )

Hi all,

I had a question about my transfections. I have the cells grown on coverslips 16 hours prior to transfection. After, I use lipofectamine to transfect my cells. At 24 hours post-transfection they look healthy and I change the media. After 48 hours, though, the cells are very streched out and starting to detach. The cells on the plastic that did not land on the coverslip, however, are just fine. I've tried using fibronectin and poly-D-lysine and still I get the same problem.

Are there any other techniques that I can try? I can see that the transfection has worked when I try to image these cells, but they're so unhealthy that I don't want to trust the data.

Yeah, that's a problem with transfecting onto glass, it just isn't quite as good for cell attachment.  One solution would be to look at the cells at an earlier time point (you say they are fine at 24 h...) or try other coating agents.  You could also move away from lipofectamine, which I find quite toxic.