cell adhesion problems in outer wells for transfeciton - (May/01/2008 )
hi all,
i'm having trouble with seeding cells in 24 or 96 well plates for transfection. each time, all the cells in the outer most wells (e.g. A1 to A6, A1 to D1, A6 to D6 and D1 to D6) either don't adhere and remain rounded, or else seem to die off after initial attachment. the inner wells don't seem to show this problem. i thought that the problem might have been with using old plates, but the problem persists even with brand new plates. we think that maybe the outer wells are getting dehydrated quickly, making the media hypertonic. but don't know for sure. i experienced this problem with NBT-II cells. have plated Vero cells today. god only knows what the outcome will be. i can't proceed much further without having this rectified.
pleeeese help.
i'm having trouble with seeding cells in 24 or 96 well plates for transfection. each time, all the cells in the outer most wells (e.g. A1 to A6, A1 to D1, A6 to D6 and D1 to D6) either don't adhere and remain rounded, or else seem to die off after initial attachment. the inner wells don't seem to show this problem. i thought that the problem might have been with using old plates, but the problem persists even with brand new plates. we think that maybe the outer wells are getting dehydrated quickly, making the media hypertonic. but don't know for sure. i experienced this problem with NBT-II cells. have plated Vero cells today. god only knows what the outcome will be. i can't proceed much further without having this rectified.
pleeeese help.
two remarks: micro titer plates are inconsistent in binding properties of wells; this is due to production processes; the chance increases that plates of the same lot have similar properties;
secondly, in deed it is often observed and described that outer wells become dry faster than inner wells; so, do not seed cells in outer wells; also, there may be problems with humidity of your incubator