3 replies to this topic

[sus12]

Hi everybody!

I want to analyze cell cycle of oxidized cells. I wonder if I have to syncronize my cells or not.
When do you syncronize the cells? I read  several articles that used syncronized cells and others used non-synchronized cells, but I do know why. When do you have to use non-syncronized and when do you have to use synchronized cells?


Thank you

[bob1]

You can use fluorescence activate cell sorting to look at the proportions of cells in different stages of the cell cycle in ordinary culture.  However, if you want to examine a particular protein you will need to synchronise so that the signal isn't confused by cells that are not at the same stage in the cell cycle.

[labrat612]

bob1, on Mar 28 2010, 08:04 PM, said:

You can use fluorescence activate cell sorting to look at the proportions of cells in different stages of the cell cycle in ordinary culture.  However, if you want to examine a particular protein you will need to synchronise so that the signal isn't confused by cells that are not at the same stage in the cell cycle.

Bob is totally correct. You can use something like nocodazole to synchronize the cells to G2/M phase and then treat your cells.

[squallweathered]

Which cells are you using?

Some cells are very easy to sync using some methods, while other cell lines will ignore the method. As standard, we usually use double thymidine blocking in our work, but for one or two lines we've needed to resort to elutriation as no chemicals would sync the cells sufficiently.

As mentioned, if there would be background signals from across the cell cycle for your particular end point then you may need to sync.