HI @dmolliver
Thanks for checking in on this.
We figured it out, but can’t be entirely sure what.
but I went back to basics. I got new, fresh laminin from Sigma.
and as you say above, instead of letting the laminin dry, we went right into wells immediately after aspirating, following this protocol Protocol for dissection and culture of murine dorsal root ganglia neurons to study neuropeptide release - PubMed
It seems to have worked. We had abundant neurons on the glass this time.
So my take away:
- Fresh reagents including PDL and laminin
- Don’t let laminin dry