Thank you very much for your protocol! I am wondering how do know where to start sectioning? Because it is a very thin piece of tissue, it would probably have been sectioned off and we couldn't tell...We tried your protocol and everything is great until when we had to sRead moreRead less
Cellular Neurobiology, Institut de Recherches Cliniques de Montreal, Canada
Thanks for your interest in our protocol. You should be able to see the retinal tissue in the OCT block when you start sectioning (it has a slightly different colour than the white OCT). I suspect you may not be sectioning the tissue in the proper orientation if you cannot see it. Make sure to rotate the OCT block by 90 degrees when you section so that you will cut across the explant (i.e. perpendicular to it). Each explant is at least 2-3 mm wide, so there should be enough tissue for you to generate many cryosections (2000 microns explant = 100 sections at 20 microns!).