Selectable light-sheet uniformity using tuned axial scanning.
Selectable light-sheet uniformity using tuned axial scanning.
复制标题
DOI：
10.1002/jemt.22795
复制
发表时间：
2017-02
影响因子：
2.5
通讯作者：
Puchalla J
中科院分区：
文献类型：
--
作者：
Duocastella M;Arnold CB;Puchalla J
光片荧光显微镜 (LSFM) 是一种光学切片技术，能够对各种样本进行快速三维 (3D) 成像，同时降低光毒性并具有出色的背景抑制能力。然而，基于椭圆形或圆形高斯光束的传统光片生成方法在光片厚度和保留该厚度的区域之间存在固有的权衡。最近，使用沿横向和光束传播方向的快速双轴高斯光束扫描证明了光片均匀性的增加。在这里，我们将类似的扫描概念应用于柱面透镜产生的椭圆光束。在这种情况下，只需要椭圆光束的 z 扫描，因此可以简化装置的实验实施。我们引入了一种简单的无量纲均匀性统计，以更好地表征扫描光片，并通过实验证明定制的均匀性比未扫描的椭圆光束高出 5 倍。该技术提供了一种简单的方法来生成和表征自定义照明轮廓，从而增强了探测器动态范围和视场的利用率，为更快、更高效的 2D 和 3D 成像打开了大门。
Light-sheet fluorescence microscopy (LSFM) is an optical sectioning technique capable of rapid three-dimensional (3D) imaging of a wide range of specimens with reduced phototoxicity and superior background rejection. However, traditional light-sheet generation approaches based on elliptical or circular Gaussian beams suffer an inherent trade-off between light-sheet thickness and area over which this thickness is preserved. Recently, an increase in light-sheet uniformity was demonstrated using rapid biaxial Gaussian beam scanning along the lateral and beam propagation directions. Here we apply a similar scanning concept to an elliptical beam generated by a cylindrical lens. In this case, only z-scanning of the elliptical beam is required and hence experimental implementation of the setup can be simplified. We introduce a simple dimensionless uniformity statistic to better characterize scanned light-sheets and experimentally demonstrate custom tailored uniformities up to a factor of 5 higher than those of un-scanned elliptical beams. This technique offers a straightforward way to generate and characterize a custom illumination profile that provides enhanced utilization of the detector dynamic range and field of view, opening the door to faster and more efficient 2D and 3D imaging.