Best way to deal with human monocytes - (Jul/03/2007 )
Does any one know the best way to deal with human monocytes on 24 well plates, as I usually get them unequally distributed, with a resultant difference between duplicate results.
I find that they are generally unequally distributed as well. The higher density being towards the periphery. If you rotate the dish in a circluar motion before they adhere you can get them to concentrate in the middle. Or if you move the dish excessively they will collect in a band as the solution sloshes about.
Another way is to let the plate lie in the hood for 5 min. This way the cells adhere quite uniformly. Most of the time, I observe that if you keep the plate immediately after seeding cells in the incubator, it redistributes the cells mostly to the periphery.
If you spin down the plate at 500rpm for 30sec it usually gently puts the cells down before they get the chance to towards the periphery.... that way i get evenly distributed cells..