Abstract: The light attenuation in biological tissues, caused by scattering and absorption, is a main issue in deep imaging. While the signal from the focal volume in multiphoton imaging is mostly generated by ballistic photons, both ballistic and scattered fluorescence photons contribute to the detected signal. The impact of emission wavelengths on deep imaging has not been investigated experimentally before. Here we perform a systematic comparison of the fluorescence attenuation in tissue at the emission wavelengths of 520 nm, 615 nm and 711 nm in three-photon imaging of mouse brain vasculature in vivo. Our results show that the impact of the emission wavelengths on multiphoton imaging depth is small compared to the excitation wavelengths.
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