I have to coat my plates with ploy-L-lysine for differentiating my cells towards neuronal lineage. I am confused whether the plate coated with PLL should be dried before seeding or it should be kept wet all the time.
Usually plates coated with PLL are dried before seeding. PLL coated, dry plates can also be stored for some time (like a month or so) at 2-8 degrees Celcius
We also dry the PLL coated cover slips or plates in 4 degrees for months and then seed on the dried ones with cell suspension, the efficiency is fine as in at least for few months the mammalian cell lines adhere well.
Absolutely agree with the two previous respondants, we dry our PLL coating before use.
Essentially we add PLL overnight at 4oC. Remove it, wash with water, and then dry at room temp overnight in a flowhood. We then wrap the plates in foil and store at room temp prior to use. My preference is always to use fresh if I can, but I know my colleagues often store the plates for 2-4weeks.
Good luck.
Hi! We do use PDL, not PLL, but I guess the procedure supposed to be similar. We use water- or ACSF-diluted PDL in concentration about 50 µg/ml, apply on the coverslip with a syringe through 0.2µ filter, let it stay about 30 min at RT, suck out the drop and let it completely dry out before cell suspension application. Works well. We culture neurons from hippocampus, if we forget them in an incubator, they may grow for month or even two - do not detach, just die 'cause of age...
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