Comparison of emission wavelengths for in vivo deep imaging of mouse brain

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.

[1]  Andreas S Tolias,et al.  In vivo three-photon imaging of activity of GCaMP6-labeled neurons deep in intact mouse brain , 2017, Nature Methods.

[2]  Q. Luo,et al.  Precise Cerebral Vascular Atlas in Stereotaxic Coordinates of Whole Mouse Brain , 2017, Front. Neuroanat..

[3]  R. Aronson,et al.  Photon diffusion coefficient in an absorbing medium. , 1999, Journal of the Optical Society of America. A, Optics, image science, and vision.

[4]  J. Ripoll,et al.  Experimental determination of photon propagation in highly absorbing and scattering media. , 2005, Journal of the Optical Society of America. A, Optics, image science, and vision.

[5]  N. Nishimura,et al.  Deep tissue multiphoton microscopy using longer wavelength excitation. , 2009, Optics express.

[6]  Bo Li,et al.  Comparing the effective attenuation lengths for long wavelength in vivo imaging of the mouse brain. , 2018, Biomedical optics express.

[7]  D. Kobat,et al.  In vivo two-photon microscopy to 1.6-mm depth in mouse cortex. , 2011, Journal of biomedical optics.

[8]  Geng Ku,et al.  Noninvasive imaging of hemoglobin concentration and oxygenation in the rat brain using high-resolution photoacoustic tomography. , 2006, Journal of biomedical optics.

[9]  K. Fujita.  [Two-photon laser scanning fluorescence microscopy]. , 2007, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme.

[10]  Yan Liu,et al.  Calibration-free quantification of absolute oxygen saturation based on the dynamics of photoacoustic signals. , 2013, Optics letters.

[11]  W. Denk,et al.  Deep tissue two-photon microscopy , 2005, Nature Methods.

[12]  A. Welch,et al.  A review of the optical properties of biological tissues , 1990 .

[13]  Frank W. Wise,et al.  In vivo three-photon microscopy of subcortical structures within an intact mouse brain , 2012, CLEO 2012.

[14]  Hao Zhang,et al.  Imaging of hemoglobin oxygen saturation variations in single vessels in vivo using photoacoustic microscopy , 2007 .

[15]  W. Denk,et al.  Two-photon imaging to a depth of 1000 microm in living brains by use of a Ti:Al2O3 regenerative amplifier. , 2003, Optics letters.