High-resolution in vivo imaging of mouse brain through the intact skull
High-resolution in vivo imaging of mouse brain through the intact skull
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DOI：
10.1073/pnas.1505939112
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发表时间：
2015-07-28
影响因子：
11.1
通讯作者：
Cui, Meng
中科院分区：
文献类型：
--
作者：
Park, Jung-Hoon;Sun, Wei;Cui, Meng
多光子显微镜是目前体内深层组织成像的首选方法。长激光波长受到的散射较少，并且 3D 限制激发允许使用散射信号光。然而，由于生物组织复杂的折射率分布，这会扰乱入射光并破坏高分辨率成像所需的光学焦点，因此成像深度仍然受到限制。在这里，我们演示了一种波前整形方案，该方案可以在扩展的校正视场 (FOV) 上通过极度混浊的生物组织（例如头骨）进行清晰成像。获得复数波前校正并以非侵入性方式直接与浑浊层共轭。使用这种技术，我们通过成年小鼠的完整头骨展示了神经树突和小胶质细胞动力学的体内亚微米分辨率成像。据我们所知，这是首次观察到完整头骨中小胶质细胞的动态形态变化，从而可以对不受外部干扰的小胶质细胞免疫活动进行真正的非侵入性研究。
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.