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The main advantage of multi-photon microscopy over conventional imaging technology is the ability to image turbid tissues in 3-dimensions. Multi-photon microscopy employs pulsed infrared lasers that generate NIR photons for non-linear excitation of UV-excitable fluorescent molecules.  Non-linear excitation requires the simultaneous absorption of two or more NIR photons that equal in energy of a single UV photon ─ this occurs in regions of a high photon flux within the sub-femtoliter focal volume of a high numerical aperture objective lens.  As a result, fluorescence outside of the focal volume is reduced during image acquisition allowing improved depth discrimination, submicron resolution, better image contrast and reduced out-of-focal volume bleaching.