Transcript: GFP Transfection Efficiency

Green Fluorescent Protein, or GFP, is widely used to study transfection efficiency and monitor intra-cellular events. The Cellometer Vision CBA offers a fast, simple, automated method of GFP analysis that requires only 20µl of cell sample.

20 µl of sample is loaded into a Cellometer Counting Chamber and inserted into the Cellometer Vision CBA Image Cytometry System. A bright field image is viewed to optimize focus and check cell morphology.

Select the GFP Transfection Assay from the drop-down menu, enter the Sample ID and Dilution Factor, then click Count. In less than 60 seconds, bright field and fluorescent cell images are captured and analyzed based on preset parameters for GFP analysis.

The total number of cells is calculated from the bright field images. Debris is excluded from cell counts based on size. GFP-positive cells are counted in the fluorescent images.

The Cellometer results table displays total cell count, cell concentration, and mean cell diameter. Data is then exported to a pre-optimized FCS Express data layout. The histogram for a control sample can be overlayed to assist with optimization of gates for treated samples. The number, concentration, and percent of GFP-negative and GFP-positive cells are displayed in the corresponding data table.

Data tables, graphs, and images can be automatically saved to a secure network and exported for presentation and publication.

Learn more about the Cellometer Vision CBA for cell-based assays at our CBA Vision page.