Measurement of Transduction and Transfection Efficiencies
Introduction of novel genetic elements into cultured cells is routinely conducted for many applications. While there are numerous methods to introduce foreign DNA into tissue culture cells, all require cell-type dependent optimization to achieve desired results. Important variables include the amount of plasmid or virus that is delivered, culture conditions, and expression kinetics of the construct. Automated imaging and analysis tools provide an efficient and robust method for measuring transfection and transduction efficiency and characterizing transient gene expression kinetics.
As shown in Figure 1, increasing amounts of GFP fusion tag lentiviral vector were added to NIH/3T3 cells in a multiwell microplate. After 24 hours, cells were imaged using the Agilent BioTek Cytation cell imaging multimode reader. The Agilent BioTek Gen5 microplate reader and imager software enabled cell counting and subpopulation analysis to report the percent transduction for each condition. Identification of total cell number can be achieved using a nuclear marker, such as Hoechst 33342 shown here, or using Gen5 label-free cell counting.
Figure 1. Transfection efficiency is measured in a multiwell microplate.