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High-resolution in vivo imaging of mouse brain through the intact skull
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High-resolution in vivo imaging of mouse brain through the intact skull
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Journal Article
High-resolution in vivo imaging of mouse brain through the intact skull
2015
Overview
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.
Significance Multiphoton microscopy has been the gold standard 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, which greatly improves the imaging depth. However, the direct application of this method to highly turbid media has been limited. Here, we present a microscope system demonstrating high resolution in vivo imaging inside highly turbid tissue. We use advanced wavefront correction with an adaptive correction plane-positioning system to compensate high-order aberrations over an extended corrected field of view. Using this technique, we demonstrate submicron-resolution imaging of neural dendrites and microglia dynamics through the intact skulls of adult mice.
Publisher
National Academy of Sciences,National Acad Sciences
Subject
/ adults
/ Animals
/ Bacterial Proteins - chemistry
/ Brain
/ Green Fluorescent Proteins - chemistry
/ Image Processing, Computer-Assisted - methods
/ Imaging, Three-Dimensional - methods
/ Luminescent Proteins - chemistry
/ Mice
/ Microscopy, Fluorescence, Multiphoton - methods
/ Nephelometry and Turbidimetry
/ Rodents
/ skull
