Hi:Thank you very much for your protocol!I am wondering how...

CL
Chris Li
Oct 15, 2018
protocol Protocol: Retinal Explant Culture
Hi:

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 s
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Michel Cayouette Author Answered Oct 16, 2018

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!).

Good luck.
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