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Proc Natl Acad Sci U S A. 2015 Jul 28;112(30):9236-41. doi: 10.1073/pnas.1505939112. Epub 2015 Jul 13.

High-resolution in vivo imaging of mouse brain through the intact skull.

Author information

1
Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA 20147.
2
Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA 20147 cuim@janelia.hhmi.org.

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.

KEYWORDS:

adaptive optics; immunology; neuroimaging; nonlinear microscopy; wavefront shaping

PMID:
26170286
PMCID:
PMC4522755
DOI:
10.1073/pnas.1505939112
[Indexed for MEDLINE]
Free PMC Article

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