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How to Quantify the Fraction of Photoactivated Fluorescent Proteins in Bulk and in Live Cells
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Capítulos
- 00:04Título
- 01:20Cell Culture and Transfection
- 02:31Imaging and Photoactivation
- 05:45Imaging Analysis and Algorithm for Ratiometric Intensity-based Quantification of Photoactivation Efficiency
- 09:09Results: Maximizing Photoactivation Efficiency
- 09:50Conclusion
Here, we present a protocol that involves genetically coupled spectrally distinct photoactivatable and fluorescent proteins. These fluorescent protein chimeras permit quantification of the PA-FP fraction that is photoactivated to be fluorescent, i.e., the photoactivation efficiency. The protocol reveals that different modes of photoactivation yield different photoactivation efficiencies.
