Multi-photon Fluorescence Microscopy
Multi-photon fluorescence microscopy is a key technology in biology. A noninvasive three-dimensional imaging at submicron scale opened new possibilities in studying of 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 few advantages like larger probing depths and in-vivo imaging.
The image brightness and its quality depend on the peak power of laser used for excitation. As the peak power is determined by pulse duration, temporal quality and repetition rate, femtosecond lasers become a key tools. Precise control of pulse duration and compression of GDD (caused by optical elements inside a microscope path) is essential to achieve shortest pulses at the sample.
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