How to seed cells evenly in small well plates - (Oct/20/2006 )
how about the suspension cells? my suspension cell has the same problem. and sometimes the cells attach to the surface and clump together to form a clone-like agglugates. what's more, as the plating is uneven, the cell counting is inaccurate too! so this is a big problem
I found out three tiny extra things can be really helpful in my practice:
1) do the rocking motion 4 times each, front to back and left to right, but avoid circular motion. (esp for dishes and large well plates)
2) add cells, put it right back to the incubator without further delay. (for small well cells)
3) make sure the shelf is leveled. if not add a piece of folded paper or something to the base to make it leveled. This is when you notice cells grwon in dishes has more cells in one side than the other.
see if pipetting up and down several times will help to get the cells loose.
I am having the same problem now. And I need some HELP If the cells are 90-100% confluent in the center and only about 50% around the well should I go ahead and split them?
Prebably yes. Cells responsed to treatment very differently at different density. You will have a hard time to reproduce the results next time.
If I re-seed the cells to a lower density will this effect the serum-starve that I am doing? This is a new and unfamiliar cell line so I am trying to test for how long I should serum starve the cells to get them to G0/G1 so if I typsinize them and re-seed them will this change anything?
Have been struggling with the same problem for 2 months now. At the end I found that the cells do not form clumps up to the point when I put them into the incubator, prior to that they're evenly distributed. The reason for aggregations turned out to be ridiculous one: the laminofore hood, placed next to the incubator, caused small shakes to the incubator, thus preventing the cells to attach whereas they are placed and causing sliding in one direction. This caused aggregates. I tried yesterday turning out the hood for several hours upon placing the cells in the incubator and found all the cells evenly distributed with hugely decreased if any aggregations.
Are the two touching each other?
A corner of the hood is touching the table where the incubator is placed.