Flow cytometry is a widely used technique for assessing transfection efficiency, providing rapid, quantitative, and single-cell resolution data. Transfection—the process of introducing nucleic acids (e.g., labeled nucleic acids)—enables researchers to determine the percentage of successfully transfected cells and evaluate transgene expression levels. This approach is critical for optimizing transfection protocols, comparing delivery methods (e.g., lipofection, electroporation), and ensuring experimental reproducibility.
Gene Expression Studies
Quantification of transfection efficiency of plasmid DNA encoding fluorescent proteins (e.g., GFP) in mammalian cells.
Monitoring the delivery and efficacy of shRNA/siRNA using reporter systems.
Viral Transfer Analysis
Assessment of lentiviral or retroviral transfer rates in target cells (e.g., CAR-T cell engineering).
Evaluation of AAV (Adeno-Associated Virus) gene transfer efficiency.
CRISPR-Cas9 Genome Editing
Measurement of CRISPR-Cas9 ribonucleoprotein (RNP) uptake through fluorescent labeling.
Sorting successfully edited cells based on reporter expression (e.g., GFP+ cells).
Therapeutic Development
Optimization of non-viral gene delivery systems (e.g., lipid nanoparticles, electroporation) for mRNA vaccines.
High-throughput screening of transfection conditions for gene therapy applications.
Generation of Stable Cell Lines
Identification and characterization of stably transfected cells using antibiotic resistance markers (e.g., puromycin selection).
Enrichment of clonal populations with high transgene expression.