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Taking Multiphoton Imaging to New Depths

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Taking Multiphoton Imaging to New Depths

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  • Author: Dr. Philip g. smith and Julien klein
TPEF microscopy was developed for live-cell imaging and uses femtosecond laser pulses for two-photon excitation of fluorophores. This excitation occurs only within a tight focal volume, where the laser’s photon density is high enough to cause two-photon absorption, eliminating out-of-focus excitation and providing inherent spatial sectioning. Because each fluorophore has a distinct two-photon excitation profile, wavelength-tunable femtosecond laser systems provide the flexibility to selectively excite the fluorophore of interest.

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