I am doing experiments with PC12 cells and TIRF microscopy. I am transfecting the cells in a 60mm dish and then culturing in 35mm dishes with a 1.0mm glass bottom, coated in PDL. I'm having difficulty with getting the cells to adhere solidly with the glass, such that the cells are flattening out which allows for TIRF microscopy. The cells do stick to the glass, but they don't strongly adhere and flatten out.
I've tried using both PDL and PLL, without any difference. I've also tried coating the glass for 20 minutes and 1 hour, washing with PBS 3 times, without any significant improvement.
I've attached some examples of the culture in flasks, in PDL coated imaging dishes, and an ideal cell under 100x TIRF magnification.
Any suggestions?
Hello, I am not a microscopy specialist but your question retains my attention because of a recent article citing use of our transfection reagent (Viromer RED) with PC12 cells. The authors used a kind of polymer-based film coating coverslips... it looks that it helps a lot for high-resolution imaging using SEM... do not know, maybe helpful for you...
Article Characterization of Cells Interactions with Patterned Azopol...
Hi
If you have used PLL and it didn't work, Use Collagen rat tail for attaching PC12 cells. Though, I have used PLL fresh evrytime to adhere my PC12 cells on dishes followed by washing!
hope it works!
I agree with Tanisha. Coating with collagen always helps when dealing with PC12 cells.
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