title: High-resolution in vivo imaging of mouse brain through the intact skull.
abstract: Multiphoton microscopy is the current method of choice for in vivo deep-tissue imaging. The long laser wavelength suffers less scattering, and the 3D-confined excitation permits the use of scattered signal light. However, the imaging depth is still limited because of the complex refractive index distribution of biological tissue, which scrambles the incident light and destroys the optical focus needed for high resolution imaging. Here, we demonstrate a wavefront-shaping scheme that allows clear imaging through extremely turbid biological tissue, such as the skull, over an extended corrected field of view (FOV). The complex wavefront correction is obtained and directly conjugated to the turbid layer in a noninvasive manner. Using this technique, we demonstrate in vivo submicron-resolution imaging of neural dendrites and microglia dynamics through the intact skulls of adult mice. This is the first observation, to our knowledge, of dynamic morphological changes of microglia through the intact skull, allowing truly noninvasive studies of microglial immune activities free from external perturbations.
authors.affiliation: Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA 20147.
authors.forename: Jung-Hoon
authors.initials: JH
authors.lastname: Park
authors.affiliation: Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA 20147.
authors.forename: Wei
authors.initials: W
authors.lastname: Sun
authors.affiliation: Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA 20147 cuim@janelia.hhmi.org.
authors.forename: Meng
authors.initials: M
authors.lastname: Cui
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry; D001921:Brain / Q000502:physiology*; D003712:Dendrites / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000737:chemistry; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008164:Luminescent Proteins / Q000737:chemistry; D051379:Mice; D017628:Microglia / Q000502:physiology; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods; D009391:Nephelometry and Turbidimetry; D059906:Neuroimaging / Q000379:methods*; D009474:Neurons / Q000502:physiology; D016011:Normal Distribution; D055095:Optics and Photonics; D019047:Phantoms, Imaging; D012542:Scattering, Radiation; D012886:Skull / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria; D049452:Green Fluorescent Proteins
keywords: adaptive optics; immunology; neuroimaging; nonlinear microscopy; wavefront shaping