I'm having a problem with transfection to synchronized cells.
My aim is to express a GFP-tagged protein and see its localization during mitosis. Actually I know It localizes to cell membrane during mitosis and I've already succeeded to see this localization. However, i need to have a high frequency of cells which are transfected and in mitosis in the same time.
I'm using lipofectamine transfection and my transfection frequency is not so bad. Also i tried double thy and single Thy+STC synchronization methods to synchronize cells and they both seem to work good as well (without transfection protocol)
However when i try to synchronize and transfect the cells, i can not have the sufficient amount of cells that are in mitosis and expressing my protein.
Any suggestions would help! Thank you!