A. Hybridization, blocking, and streptavidin/antibody incubations are all performed with a top coverslip overtop the fiber coverslip. To ensure the solution has been evenly wicked across the fiber coverslip, solutions are added to a clean coverslip and, by holding the bottom of the slide, the fiber coverslip is tapped on top.
B. Sealing the fiber coverslip and top coverslip with rubber cement allows hybridization to be performed on a slide warmer rather than in a humid chamber. Use fresh rubber cement and cut a centimeter off the tip of a 1000 μl pipette tip for ease of pipetting. Excess rubber cement is not detrimental, but only a fine ribbon of cement is needed to completely seal the coverslips.