Counting of dissociated DRG cultures for consistent assay plating

What are the best ways to count dissociated DRG neurons before plating them?
There is a lot of debris in an acute suspension of DRGs so it’s difficult to make out neurons from debris and non-neuronal cells.

Hemocytometers works well for clean suspensions but are difficult for messy suspensions like DRG. I’ve used live dyes like calcein AM before and it helps to pick out cells.

@dmolliver @tberta @thicunha

Does your protocol use a 70 um cell strainer followed by a 15% BSA cushion? I find most debris is gone, and the neurons are nice and round so are easy to count. If useful, happy to share my protocol.

Thanks @adgaudet. I would love to see your protocol. We don’t do the BSA cushion, although I know some do. I actually want non-neuronal cells in my cultures, but it would be good to remove them if needed, and debris. a.chamessian@wustl.edu. Or if you’re so inclined, consider adding to the community protocols at Pain Researcher Community Protocols - research group on protocols.io