Hi all,

I am attempting to image a mouse brain slice (~200um) on a dish [picture] and then stain it with a secondary antibody. After the staining I'd like to image it again. My problem is that to avoid/mitigate an image registration nightmare I'd like to learn of some technique or protocol to either bind or fix my tissue in place so it won't move during the staining process (that involves blocking steps, washing, & using an orbital shaker).

The dish has a smaller circle in the middle which acts as a well. There's a smaller square cover glass under the dish and a bigger one on top of the well. I apply "nail polish top coat" to opposite corners of the top cover glass to fix the cover glass in place and stop the brain tissue from moving.

Thank you for any helpful insight