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two-photon excited fluorescence: Two-photon excited fluorescence (TPEF) is a nonlinear optical method that allows imaging of biological cells and living tissue. The advantage of TPEF in comparison to conventional fluorescence microscopy is that it provides natural confocality and allows sectioning of the sample. Because it typically uses near-infrared excitation light, the penetration depth is significantly increased. TPEF is implemented as fast imaging microscopy for noninvasive optical pathology. TPEF has been used in numerous fields including physiology, neurobiology, embryology and tissue engineering.

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