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Five-Color STED With a Single Depletion Laser and Fluorescence Lifetime Phasor Separation
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Dive into the innovative world of STED microscopy and explore how researchers are pushing the boundaries of super-resolution imaging in this episode of Listen In.
Traditional STED microscopy achieves high resolution by using a doughnut-shaped depletion laser to selectively suppress fluorescence, but multicolor experiments often face challenges such as laser misalignment and dye bleaching.
Explore the game-changing technique of using a single depletion wavelength combined with phasor-based FLIM separation. This method allows scientists to distinguish multiple fluorophores with similar emission spectra but different fluorescent lifetimes, significantly increasing the number of usable spectral channels.
Discover how researchers successfully depleted five fluorochromes across different channels using a single laser and utilized the phasor plot for quick and easy visualization of staining quality and lifetime analysis. This approach not only doubles the number of distinguishable colors in laser scanning microscopy but also simplifies the process, making it accessible to many scientists with minimal training.
Watch the full presentation here: https://microscopyfocus.com/five-color-sted/
Browse all episodes of the Listen In Series here: https://listen-in.bitesizebio.com/
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