Assessing the imaging performance of light sheet microscopies in highly scattering tissues
Assessing the imaging performance of light sheet microscopies in highly scattering tissues
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DOI：
10.1364/boe.7.000454
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发表时间：
2016-02-01
影响因子：
3.4
通讯作者：
Liu, J. T. C.
中科院分区：
文献类型：
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作者：
Glaser, A. K.;Wang, Y.;Liu, J. T. C.
光片显微镜（LSM）已成为一种光学成像方法，用于对相对透明的样品进行高时空体积成像。虽然这种能力使得该技术在发育生物学等领域具有高度影响力，但涉及高度散射厚组织的应用在很大程度上尚未被探索。在这里，我们采用蒙特卡罗模拟来探索 LSM 在浑浊介质成像中的应用。特别是，由于其与双轴共焦 (DAC) 显微镜的相似性，我们将 LSM 性能与点扫描 (PS-DAC) 和线扫描 (LS-DAC) 双轴共焦显微镜技术进行比较，这些技术先前已被证明可以在分别类似于 9 - 10 和 3 - 4 的往返光学长度下产生高质量图像。这项研究的结果表明，使用宽场收集 (WF-LSM) 的 LSM 在厚组织中提供了与 LS-DAC 相当的性能，因为它们都利用聚焦在一维（即线或片）的照明光束。另一方面，使用共焦线检测 (CL-LSM) 的 LSM 更类似于 PS-DAC 显微镜，其中照明光束在二维上聚焦到一个点。由于使用较低数值孔径 (NA) 照明光束沿照明轴扩展成像，LSM 的成像深度仅略逊于 DAC（分别类似于 WF-LSM 和 CL-LSM 的 2 - 3 光学长度和 6 - 7 光学长度）。因此，我们得出的结论是，深度成像的能力主要取决于显微镜技术的共焦性。此外，我们发现成像分辨率主要取决于采集数值孔径，并且在均匀散射介质中对成像深度相对不变。我们的结果表明，使用光片显微镜对高度散射组织进行表面成像是可能的。 (C) 2016年美国光学学会
Light sheet microscopy (LSM) has emerged as an optical-imaging method for high spatiotemporal volumetric imaging of relatively transparent samples. While this capability has allowed the technique to be highly impactful in fields such as developmental biology, applications involving highly scattering thick tissues have been largely unexplored. Herein, we employ Monte Carlo simulations to explore the use of LSM for imaging turbid media. In particular, due to its similarity to dual-axis confocal (DAC) microscopy, we compare LSM performance to point-scanned (PS-DAC) and line-scanned (LS-DAC) dual-axis confocal microscopy techniques that have been previously shown to produce high-quality images at round-trip optical lengths of similar to 9 - 10 and similar to 3 - 4 respectively. The results of this study indicate that LSM using widefield collection (WF-LSM) provides comparable performance to LS-DAC in thick tissues, due to the fact that they both utilize an illumination beam focused in one dimension (i.e. a line or sheet). On the other hand, LSM using confocal line detection (CL-LSM) is more analogous to PS-DAC microscopy, in which the illumination beam is focused in two dimensions to a point. The imaging depth of LSM is only slightly inferior to DAC (similar to 2 - 3 and similar to 6 - 7 optical lengths for WF-LSM and CL-LSM respectively) due to the use of a lower numerical aperture (NA) illumination beam for extended imaging along the illumination axis. Therefore, we conclude that the ability to image deeply is dictated most by the confocality of the microscope technique. In addition, we find that imaging resolution is mostly dependent on the collection NA, and is relatively invariant to imaging depth in a homogeneous scattering medium. Our results indicate that superficial imaging of highly scattering tissues using light sheet microscopy is possible. (C) 2016 Optical Society of America