Multi-photon Fluorescence Microscopy

Multi-photon fluorescence microscopy is a key technology in biology. Noninvasive three-dimensional imaging at the submicron scale opened new possibilities in studying tissues.

The mechanism is based on multi-photon excitation of fluorophores, usually in near-infrared wavelength. Relaxing fluorophore emits a photon in visible range which is detected. However, the nonlinear character of the process gives a few advantages like larger probing depths and in-vivo imaging.

The image brightness and its quality depend on the peak power of the laser used for excitation. As the peak power is determined by pulse duration, temporal quality, and repetition rate, femtosecond lasers become a key tool. Precise control of pulse duration and compression of GDD (caused by optical elements inside a microscope path) is essential to achieve the shortest pulses at the sample.

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