Automated Optimization of Cell Transfection

Cell transfection is an essential technique for interrogating cellular pathways. Determining the optimal conditions for introducing nucleic acids into cells can be a time consuming endeavor at the initiation of a cell biology experiment or screen. Here we demonstrate how the Biomek FXP Workstation can be used to identify the optimal conditions for siRNA transfection in a single experiment. Utilizing the enhanced multichannel selective tip pipetting, factorial combinations of transfection reagents and concentrations, cell number, and fluorescent oligonucleotide concentrations were plated in 384-well plates. This higher density plate format conserved reagents while also facilitating the replicate wells necessary to determine variability within a given condition. 24 hours after transfection, cells were stained with Draq7 and transfection efficiency and cell toxicity were measured on the SpectraMax i3 Multi-Mode Detection Platform with MiniMax 300 Imaging Cytometer. Automation enabled the miniaturization and execution of a complex optimization experiment, thereby reducing reagent costs and accelerating the timeline to meaningful transfection assays and screens.