Transfection Optimization Using Expression Analysis

Optimization of cell transfection typically includes determining the optimum mass of plasmid to be transfected and the time point after transfection that provides the maximum expression. Too much plasmid can be toxic and lead to compromised cell growth; too little plasmid will reduce the overall transfection efficiency. In this application note it is shown the Celigo cytometer from Brooks Life Science Systems enables cell-based fluorescence quantification in a variety of multi-well formats and flasks without the requirement for counterstains. Cells can be left undisturbed in the plate or flask, imaged within 20-30 minutes, depending on the format, and then returned to the incubator for continued culturing. This facilitates time-course studies, such as monitoring fluorescence after a transient transfection, dramatically reducing the effort and time to perform the experiments.