多维数字扫描光片显微镜可实现均匀的荧光激发和对比度增强成像。
Multidirectional digital scanned light-sheet microscopy enables uniform fluorescence excitation and contrast-enhanced imaging.
机构信息
Department of Mechanical Engineering, University of Washington, Seattle, WA, USA.
Department of Pathology, University of Washington, Seattle, WA, USA.
出版信息
Sci Rep. 2018 Sep 17;8(1):13878. doi: 10.1038/s41598-018-32367-5.
Light-sheet fluorescence microscopy (LSFM) has emerged as a powerful method for rapid and optically efficient 3D microscopy. Initial LSFM designs utilized a static sheet of light, termed selective plane illumination microscopy (SPIM), which exhibited shadowing artifacts and deteriorated contrast due to light scattering. These issues have been addressed, in part, by multidirectional selective plane illumination microscopy (mSPIM), in which rotation of the light sheet is used to mitigate shadowing artifacts, and digital scanned light-sheet microscopy (DSLM), in which confocal line detection is used to reject scattered light. Here we present a simple and passive multidirectional digital scanned light-sheet microscopy (mDSLM) architecture that combines the benefits of mSPIM and DSLM. By utilizing an elliptical Gaussian beam with increased angular diversity in the imaging plane, mDSLM provides mitigation of shadowing artifacts and contrast-enhanced imaging of fluorescently labeled samples.
光片荧光显微镜（LSFM）已经成为一种快速、高效的 3D 显微镜方法。最初的 LSFM 设计利用了静态光片，称为选择平面照明显微镜（SPIM），由于光散射，它表现出阴影伪影和对比度降低的问题。这些问题在一定程度上得到了解决，例如多方向选择平面照明显微镜（mSPIM），其中光片的旋转用于减轻阴影伪影，以及数字扫描光片显微镜（DSLM），其中使用共聚焦线检测来拒绝散射光。在这里，我们提出了一种简单而被动的多方向数字扫描光片显微镜（mDSLM）架构，它结合了 mSPIM 和 DSLM 的优点。通过利用在成像平面中具有增加的角多样性的椭圆高斯光束，mDSLM 提供了对阴影伪影的缓解和对荧光标记样品的对比度增强成像。
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