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Two-photon excitation microscopy

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Two-photon excitation microscopy (TPEF or 2PEF) is a fluorescence imaging technique that is particularly well-suited to image scattering living tissue of up to about one millimeter in thickness. Unlike traditional fluorescence microscopy, where the excitation wavelength is shorter than the emission wavelength, two-photon excitation requires simultaneous excitation by two photons with longer wavelength than the emitted light. The laser is focused onto a specific location in the tissue and scanned across the sample to sequentially produce the image. Due to the non-linearity of two-photon excitation, mainly fluorophores in the micrometer-sized focus of the laser beam are excited, which results in the spatial resolution of the image. This contrasts with confocal microscopy, where the spatial resolution is produced by the interaction of excitation focus and the confined detection with a pinhole.

Two-photon excitation microscopy - Wikipedia Jump to content From Wikipedia, the free encyclopedia Fluorescence imaging technique Not to be confused with second-harmonic imaging microscopy . Volumetric two-photon calcium imaging of 852 neurons in awake mouse brain. The imaged volume spans 1400×1900×460µm³, sampled at 30 volumes per second [ 1 ] . Volumetric two-photon imaging of FITC -labeled microvessels in awake mouse brain. The depth-color-coded imaged volume spans 540×550×510µm³, sampled at 30 volumes per second [ 1 ] . Two-photon excitation microscopy of mouse intestine . Red: actin . Gre

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