Has anyone had any success sorting neurons and astrocytes? Preferably not with transgenic lines, but rather with antibodies. Any help is appreciated.
Hi @ShanTan we have established sorting of astrocytes in the spinal cord of mice using anti-ACSA2 antibody anti-ACSA2 - PE (IH3-18A3, Miltenyi Biotec, 1:250). Neurons are more complicated due to viability. You can find more information in our preprint paper (https://www.biorxiv.org/content/10.1101/2021.01.25.428142v2).
Where are you dissociating neurons from - brain or spinal cord? Adult spinal cord neurons are very hard to dissociate and keep alive. You could try the following:
I messed around with the NeuO dye. It’s not as selective as I wanted. Bu those are options I’m aware of. You could also possibly try NeuroTrace. I’ve never used it on dissociated cells but it may work.
Thanks @thicunha! Will definitely give your protocol a try.
@achamess I’d be attempting to sort neurons from spinal cord tissue. Do you think it’s better to get a transgenic line to label neurons? I guess the labelling of the neurons themselves isn’t the issue, it’s keeping them alive and getting enough of them from tissue to be able to do anything?
This paper uses transgenic mice to select spinal neurons. They used adolescent mice. and they had to sort and select because neuron viability is an issue. Spinal neurons are hard to extract from all their milieu without beating them up. But it’s possible.