I usually coated the coverslips with PLL with conc 1 % and the DRGs neurons did not attach well to coverslips and I do not know why. Also , if somebody know a method for fixation of this neurons or any neurnal cells generally , I usually use 4% paraformaldehyde and after fixation I foundsome cells leave coverslips and some damage. so if somebody dealing with these cells .please, help me
Hello!
To coat DRGs I used a solution of 0,2 mg/ml Laminin in HBSS and 0,02 mg/ml PDL in HBSS. Coverslips with 100μl of coating solution were stored at 4 C for 2 hours before use. Right before use I washed them with dH2O.
I strongly recommend this protocol https://www.nature.com/articles/nprot.2006.461
As for fixation, I used 4% PFA for 15 minutes and i washed coverslips with PBS.
Konstantina Dimoula Thanks so much for this valuable suggestions. I will try this super protocol. I think the problem in coating of cells was not good. Thanks again
coat with poly-D-lysine hydrobromide
See how patch clamp reserachers do it. Look into: Van Duijn B, Ince C et al., (1992) Whole-cell patch clamp experiments: a way to introduce substances into cels. An overview of potential difficulties in plant and animal cells: In: Biotechnology applications of microinjections, microscopic imaging and fluorescence. Ed P.H.Bach pp001-010 Plenum Pub Co Ny USA
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