Applications

Transfection Efficiency

Monitor and quantify the efficiency and timecourse of gene transfection using GFP/RFP constructs.


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Key features

Key features

Reagent Optimization

Using GFP/RFP tagged constructs, easily compare the efficiencies and time-course of expression with different transfection conditions e.g. cationic lipid reagents, amount of DNA, exposure time and presence of serum

Amenable to multiple tissue culture vessels

Use either T-flasks, microplates or dishes. Low though to high magnification (4x, 10x, 20x). Image multiple areas of your vessel to highlight spatial variations and optimize cell distribution

Monitor Cell Confluence and Cell Health

Prior to transfection, accurately assess cell confluence to standardize cell transfection technique. Post transfection, monitor the effect of reagents on cell health using label-free phase segmentation

Monitor Expression Intensity

Use histograms to monitor the distribution of GFP/RFP intensity of transfected cells

Example data

Example data

Upper left: Time-course of nuclear GPF expression in A549 human lung carcinoma cells transduced with IncuCyte® NucLight Green lenti- virus. Abscissa: time (h), ordinate: fluorescent object count (per mm2). Upper right: IncuCyte® blended HD-phase contrast and green fluorescence image of A549 cells transduced with NucLight Green lenti-virus (48h). Note nuclear restriction and homogenous expression of GFP. Transduction of cells with NucLight Green is titratable (lower left: A549 cells, lower right: HUVECs) and transduction efficiencies can be enhanced with Polybrene (lower left: A549 cells).

Time-course of nuclear GPF expression in A549 human lung carcinoma cells transduced with IncuCyte® NucLight Green lenti- virus

IncuCyte® blended HD-phase contrast and green fluorescence image of A549 cells transduced with NucLight Green lenti-virus

Note nuclear restriction and homogenous expression of GFP. Transduction of cells with NucLight Green is titratable

transduction efficiencies can be enhanced with Polybrene

Technical Resources

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