Monitor Transfection Efficiency With the Incucyte® Live-Cell Analysis System

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

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4th Edition! Live-Cell Analysis Handbook

A guide to real-time live-cell imaging & analysis

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cytokine app note

Application Note

Gain additional insights into the mechanism of immune cell killing by combining live cell analysis and flow cytometry into a single workflow.

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Key Advantages

Key Advantages

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 GFP expression in A549 human lung carcinoma cells transduced with IncuCyte® NucLight Green lentivirus. 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 lentivirus (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).

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