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. As cells proliferate, the number of transfected cells will increase at the expense of diluting the plasmid in each transfected cell. Thus there will be an optimum time point for maximum number of expressing cells.

The Celigo adherent cell cytometer is a benchtop in situ cellular analysis system that rapidly provides high integrity whole well
images for routine brightfield and fluorescent cellular analysis.
The Celigo cytometer provides brightfield and three channels of
fluorescence capabilities for visualizing and quantifying cellular
responses in 1,536- to 6-well plates and flasks. The system provides full resolution at 1 µm/pixel (Figure 1). Developed for brightfield and fluorescence imaging, it incorporates proprietary optics and software to image and identify cells in brightfield with consistent contrast across the whole well all the way up to the well edge. This enables the ability to use the brightfield image for defining the position and area of the cells without the need to add additional fluorescent probes.