The ability to visualize deep brain structures in vivo with high spatial resolution is of rising interest to investigate neuronal physiology and cerebral vasculature. Optical imaging offers non-invasive, high-resolution in vivo microscopy techniques to observe brain tissue and its surrounding environment. Two-photon fluorescence laser- scanning microscopy (2PM) can overcome depth limitations by using nonlinear excitation. The ideal approach for deep imaging in brain is to use both high energy pulses and longer excitation wavelengths.
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