I need to stick HCT116 cells from suspension to coverslips for immunofluorescence. I am doing short time points so I cannot have the cells sitting on the coverslips for 20-30 minutes, I plan to fix them with formaldehyde at each time point, wash out the Formaldehyde and then find a way to get them onto coverslips. I've heard poly-L-lysine and concanavilin will not work with fixed cells. I've heard about CyGel but I'm afraid of it melting since it melts below 23 degrees C (per the manual). I've heard good things about Cell-Tak from corning. Does anyone have experience with this or any other methods - these are all the ones I am aware of.
I had experience in culturing epithelial cells grow in suspension. To do immunofluorescence, I usually cytospin them onto a poly-L-lysine coated glass slide and then proceed with the immunofluorescence protocol. Cytospinning is essential to help sticking the cells onto poly-L-lysine by centrifugal force. By the way, I used paraformaldehyde to fix the cells.
If you want to use this method, the problem lies in whether you have a cytospin centrifuge and whether using coverslips is an important part of your experiment because the cytospin I know of only fits glass slide.
A lot of people are afraid of the morphological changes that occur during cytospin. I would plate them onto coverslips live and let them naturally flatten, but they take so long it is impossible to do time points - has anyone tried poly-L-lysine coating coverslips or any other surface to see if the cells will attach and flatten out faster?
Hi David, to perform IF stainings with suspension cells you can simply resuspend the cells in PBS1x, and let the cells to adhere on a coverslip for about 15 min at 37C. It works very nicely!