I want to perform immunocytochemistry on transfected HEK cells. I’ve been getting them to grow and adhere to Ibidi 12 well glass slides while they’re growing. I can do media changes and they stay. But once I take the out and fix and then do some washes, they are mostly gone. it’s frustrating. The glass is coated with PDL for 2 hours before seeding the cells.
Does anyone have tricks for getting HEK cells to stay on the glass for ICC?
Indeed, some cells very weakly adhere to the glass surface. However in my hands after fixation, things don’t get worse. I used glass from the various manufacturer and coated it with 50 ug/mL atelocollagen for 1 or 2 h. For fixation, I used 4% formaldehyde.
Hope it helps.
Generally, HEKs are a bit troublesome when it comes to sticking. We use 0.01% Poly-L-lysine (freshly diluted stock) for 30min-1hr before seeding cells. When I have seen poor adherence it is mainly because of the quality of poly-l-lysine use (either not freshly diluted or old stock).