Direct visualization of microvasculature provides significant insights in microcirculation and critically impacts the diagnosis and treatment of microcirculatory diseases. Recently, we discovered that the high-energy Soret fluorescence of hemoglobin peaked at with an extremely short lifetime becomes strongly visible under two-photon excitation. Based on the distinct spectral and temporal characteristics of hemoglobin fluorescence, we demonstrated that two-photon fluorescence microscopy could become a powerful tool for label-free in vivo imaging of microvasculature in tissue.
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