Christine Jolicoeur Cellular Neurobiology, Institut de Recherches Cliniques Montreal, Canada

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Michel Cayouette Cellular Neurobiology, Institut de Recherches Cliniques de Montreal, Canada
2 protocols

Christine Jolicoeur

Cellular Neurobiology, Institut de Recherches Cliniques de Montreal, Canada

Research focus

Neuroscience

2 Author merit
Reviewer merit
Editor merit

2 Protocols published

Dissociated Retinal Cell Culture

Authors: Christine Jolicoeur and Michel Cayouette, date: 01/20/2014, view: 13881, Q&A: 0

The retina is a relatively simple and accessible part of the central nervous system, making it a powerful model to study cell fate specification mechanisms. Multipotent retinal progenitor cells (RPCs) give rise to seven major classes of retinal cell ...

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Retinal Explant Culture

Authors: Christine Jolicoeur and Michel Cayouette, date: 01/20/2014, view: 19509, Q&A: 3

A particularly powerful culture method for the retina is the explant assay, which consists in culturing a small piece of retina on an organotypic filter. Retinal explants can be prepared any time between embryonic day 13 (E13) and postnatal day 4 ...

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maryam hejazi

university of alberta

Hi ,

I used your protocol to make retina explant everything went well, until PFA fixation, where I tried to cover my retina sections with PFA, and added PFA to the membrane very softly, but my explant detached and got floated in the PFA solution!

11/21/2018 1:41:15 PM Reply

maryam hejazi

university of alberta

Thank you, I am starting new explant tomorrow, hopefully work this time:)

2018-11-22 08:11:46

Christine Jolicoeur

Cellular Neurobiology, Institut de Recherches Cliniques de Montreal, Canada

Hi Maryam, sometimes there is some contamination that can be visible around the explants. It looks like a white ring. The explants still grow and this contamination doesn't take over but it is not optimal to grow explants in these conditions. For the medium, no need to change the composition but make sure you change it twice a week. The evaporation of the medium is quit substential and it depend on the kind of incubator you have. We have an incubator the monitor the humidity to 90% and we calculated that 30 microliters of medium evaporate every day. The protocol says that 80% of the medium should be change but you can change the totality and it won't affect your culture. Hope it helps! Good luck, Christine

2018-11-21 18:04:29

maryam hejazi

university of alberta

Thanks Christine, I will make sure to follow these steps next time. What are the reasons that my culture might be unhealthy? Do I have to switch the media to different one?

Thanks

2018-11-21 14:03:04

Christine Jolicoeur

Cellular Neurobiology, Institut de Recherches Cliniques de Montreal, Canada

Hi Maryam,
This can happen. Make sure you are using room temperature PFA (very important). You should first remove the medium from under the filter and replace it with PFA. Fix for 5 minutes and than add PFA on top of the explants. If your explants still detach easily it is often a sign that they are not very healthy.

2018-11-21 13:52:21