Transfection Efficiency - Actin and Integrin Double Transfection
HeLa cells transfected with integrin (orange/TRITC) and actin (red/Cy5) plasmids, with additional DAPI (blue) nuclear stain and wheat germ agglutinin (green/FITC) membrane stain. This dataset demonstrates how to determine transfection efficiency for both single and double plasmid transfections. Shows relatively low transfection efficiency, highlighting the value of Cytely for method optimization.
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Methodology
HeLa cells transfected with integrin and actin plasmids. Four-channel fluorescence imaging: DAPI (blue, nuclei), FITC/wheat germ agglutinin (green, membrane), TRITC/integrin (orange), Cy5/actin (red).
Usage examples
Quantifying single-plasmid transfection efficiency. Identifying double-positive cells expressing both plasmids. Optimizing transfection protocols. Multi-parameter fluorescence analysis.
Technical notes
Low transfection efficiency makes this an excellent dataset for demonstrating the importance of protocol optimization. Sequential gating allows precise identification of single-positive and double-positive populations.
Assays in this dataset
Cell Health & Morphology
Transfection Efficiency
Quantifies the fraction of cells expressing one or more transfected constructs.
Cell Health & Morphology
Cell Counting & Viability
Counts total cells and classifies them as live or dead based on marker intensity, providing the most fundamental readout for toxicity and dose-response experiments.
Cell Health & Morphology
Cell Morphology & Phenotypic Profiling
Measures the size, shape, and fluorescence intensity of individual cells across multiple markers.
Other
Sample Purity & Quality Control
Identification and quantification of contaminants in cell preparations
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