trouble to grow cells on top of coverslip
Posted 11 September 2009 - 12:40 PM
I used chamber slides but couldn't get the same result when I prep cells on 12 well plate....agrrrrr
Should I glue coverslip with vaseline or whatever? Please help!!!!!!!!!
Posted 11 September 2009 - 01:25 PM
However, I normally submit the cover slips to UV light in the hood to sterilize and then seed.
I don't think you need to HCL wash the slips, this may change the charge of the cover lsips and lead to less binding.
I put the slip into a 24 well plate add media and THEN add the cells. This order seems to work well.
Also, using fresh media makes a difference.
Posted 11 September 2009 - 01:49 PM
Posted 11 September 2009 - 02:12 PM
Solution will be how to prevent cells get in between coverslip and culture dish. I'll try the not-to-spill-over method, wish me good luck! (I have coworkers tend to slam the incubater door...) Thank you!!!
Posted 14 September 2009 - 02:02 AM
As for the sterilizing - I use 70% ethanol and leave to dry. Never had a problem before.
Posted 15 September 2009 - 06:32 PM
- use the lid of a 35mm dish to hold about 1ml of 2% gelatin solution
- take your autoclaved coverslips and dip them into the gelatin solution (do both sides)
- take the coated coverslip and place it inside another 35mm dish
- close the dish and let the gelatin dry (I usually turn on the UV and let it dry overnight)
- repeat the above steps to make as many dishes as you need (I usually make stocks)
In this method, the coverslip is adhered to the bottom when you initially seed the cells, so this prevents the majority of cells from sneaking to the bottom. Don't worry about the coverslip being stuck, once the coverslip is submerged in media, it will eventually be "unstuck".
Edited by Bill Nye, 15 September 2009 - 06:34 PM.
Posted 16 September 2009 - 09:24 AM