I presume you are coating c/s o/n and drying to get good adherence. You could try precoating c/s with polylysine beforehand - often used to coat plastic t/c plates.

I have not tried UV fixation or derivitisation of glass prior to collagen coating, but these could be possibilities

I am attempting to grow explants on Millipore Isopore membrane filters placed on gelatin gel-foam supports. I presume it is possible to coat the membranes with collagen before placing explants on top, but have not tried it (without the gel foam). It would have more limited visibility compared to glass. Glass, unfortunately, is often not a particularly good surface for adherence.

Would be interested to know how you solve this problem.

Ian

Thank you very much!

Yes, I know about the problems with coverslips, regarding adherence. Yet, better to use coverslips than multiwell slides, that often give autofluorescence.

I tried to coat the coverslips with gelatin prior to placing the collagen drop, but it did not seem to help. Today I am trying a collagen coating before placing the gel and as soon as I have PLL-coated coverslips I will try on those. I will let you know how I'm getting on and if I can solve my problem.

THANKS again!

Hello. I coat coverslips in collagen successfully all of the time. I start by collagen coating the night before. I place the collagen on the coverslip and leave it in the t/c hood with the UV on overnight. The UV helps the collagen cross-link. If you don't have overnight, you can always coat just a couple hours before. When the slip looks dry the coverslip is ready to go. The coverslips I use are from Fisher: 12-545-81. Also, it might help to acid wash the coverslip before you start: Just swirl in a 3:1 solution Nitric acid:HCl. Then rinse with water, 95% EtOh, 70% EtOH, 95% EtOH and flame dry. After this you can start with the collagen. Hope this helps.