Opt Express. 2020 May 11;28(10):15587-15600. doi: 10.1364/OE.393728.
Light-sheet fluorescence microscopy (LSFM) helps investigate small structures in developing cells and tissue for three-dimensional localization microscopy and large-field brain imaging in neuroscience. Lattice light-sheet microscopy is a recent development with great potential to improve axial resolution and usable field sizes, thus improving imaging speed. In contrast to the commonly employed Gaussian beams for light-sheet generation in conventional LSFM, in lattice light-sheet microscopy an array of low diverging Bessel beams with a suppressed side lobe structure is used. We developed a facile elementary lattice light-sheet microscope using a micro-fabricated fixed ring mask for lattice light-sheet generation. In our setup, optical hardware elements enable a stable and simple illumination path without the need for spatial light modulators. This setup, in combination with long-working distance objectives and the possibility for simultaneous dual-color imaging, provides optimal conditions for imaging extended optically cleared tissue samples. We here present experimental data of fluorescently stained neurons and neurites from mouse hippocampus following tissue expansion and demonstrate the high homogeneous resolution throughout the entire imaged volume. Utilizing our purpose-built lattice light-sheet microscope, we reached a homogeneous excitation and an axial resolution of 1.2 µm over a field of view of (333 µm).
光片荧光显微镜（LSFM）有助于对发育中的细胞和组织中的小结构进行三维定位显微镜检查和神经科学中的大视场脑成像。晶格光片显微镜是一种具有很大潜力的新技术，可以提高轴向分辨率和可用的视场尺寸，从而提高成像速度。与传统 LSFM 中用于光片产生的常用高斯光束相比，在晶格光片显微镜中，使用具有抑制旁瓣结构的低发散贝塞尔光束阵列。我们使用微制造的固定环掩模开发了一种简便的基本晶格光片显微镜，用于晶格光片的产生。在我们的设置中，光学硬件元件可实现稳定且简单的照明路径，而无需空间光调制器。这种设置与长工作距离物镜以及同时进行双色成像的可能性相结合，为成像扩展的光学透明组织样本提供了最佳条件。我们在此展示了经组织扩张后荧光染色的神经元和神经突的实验数据，并证明了整个成像体积内具有高度均匀的分辨率。利用我们专门设计的晶格光片显微镜，我们实现了在（333 µm）视场范围内的均匀激发和 1.2 µm 的轴向分辨率。
