Saturated two-photon excitation fluorescence microscopy for the visualization of cerebral neural networks at millimeters deep depth
Journal
Journal of biophotonics
Journal Volume
12
Journal Issue
1
Date Issued
2019
Author(s)
Abstract
Optical imaging is a key modality for observing biological specimen with higher spatial resolution. However, scattering and absorption of light in tissues are inherent barriers in maximizing imaging depth in biological tissues. To achieve this goal, use of light at near-infrared spectrum can improve the present situation. Here, the capability of saturated two-photon saturated excitation (TP-SAX) fluorescence microscopy to image at depths of >2.0 mm, with submicron resolution in transparent mouse brain imaging, is demonstrated. At such depths with scattering-enlarged point spread function (PSF), we find that TP-SAX is capable to provide spatial resolution improvement compared to its corresponding TPFM, which is on the other hand already providing a much improved resolution compared with single-photon confocal fluorescence microscopy. With the capability to further improve spatial resolution at such deep depth with scattering-enlarged PSF, TP-SAX can be used for exquisite visualization of delicate cerebral neural structure in the scattering regime with a submicron spatial resolution inside intact mouse brain. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Subjects
brain imaging; nonlinear microscopy; saturated fluorescence excitation microscopy; two-photon fluorescence microscopy
SDGs
Other Subjects
Brain mapping; Fluorescence; Fluorescence microscopy; Histology; Image resolution; Infrared devices; Light; Mammals; Near infrared spectroscopy; Optical transfer function; Particle beams; Photons; Tissue; Visualization; Brain imaging; Confocal fluorescence microscopy; Fluorescence excitation; Nonlinear microscopy; Scattering and absorption; Submicron spatial resolution; Two-photon excitation fluorescence microscopy; Two-photon fluorescence microscopy; Brain; carbocyanine; cyanine dye 3; animal; brain; C57BL mouse; diagnostic imaging; light related phenomena; metabolism; mouse; multiphoton microscopy; nerve cell network; procedures; Animals; Brain; Carbocyanines; Mice; Mice, Inbred C57BL; Microscopy, Fluorescence, Multiphoton; Nerve Net; Optical Phenomena
Publisher
WILEY-V C H VERLAG GMBH
Type
journal article