Lattice light sheet microscope for fast dual-color long-term imaging

许多重要的生理过程，如发育和再生、生物节律和衰老，都是在多个时间尺度上调控和表现的。对这些过程的研究需要在多个层面上进行测量，从生物体和组织行为到细胞和分子事件。因此，为了理解复杂的生理过程，需要能够在长时间内以高时间和空间分辨率对这些过程进行成像的方法。在长时间内以高空间和时间分辨率成像生物过程的一个主要挑战是大多数高分辨率光学切片成像方法引起的光毒性。光片显微镜提供了独特的特点，特别照亮当前成像焦平面，从而导致一个更温和的照明标本。此外，点阵光片技术的出现使光片成像在亚细胞分辨率。对于德累斯顿工业大学的许多研究项目来说，一个强大的晶格光片成像平台是必不可少的，它允许对光敏生物标本进行调查。在这里，我们申请了一个点阵光片平台，可以在很长一段时间内进行快速和温和的多色实验。晶格光片技术首次允许对快速生物过程进行亚细胞分辨率的长期多色成像。因此，这项技术将成为研究许多高度相关的生物过程的基础。该平台将建立在德累斯顿工业大学分子和细胞生物工程中心（CMCB）的光学显微镜设施（LMF）中，从而确保所有用户的开放访问和最佳支持。

Many important physiological processes such as development and regeneration, biological rhythms, and aging are regulated and manifest at multiple temporal scales. The investigation of these processes requires measurements at multiple levels, ranging from organism and tissue behavior to cellular and molecular events. Hence, to understand complex physiological processes, methodologies are needed that can image these processes with a high temporal and spatial resolution over long time periods. A major challenge in imaging biological processes with high spatial and temporal resolution over extended time periods has been the phototoxicity that is caused by most high-resolution optical sectioning imaging approaches. Light sheet microscopy offers the unique feature of specifically illuminating the currently imaged focal plane, thus resulting in a much gentler illumination of the specimen. Moreover, the advent of the lattice light sheet technology has enabled light sheet imaging at subcellular resolution. A robust lattice light sheet imaging platform allowing for the investigation of light-sensitive biological specimen is essential for many research projects within TU Dresden. Here, we apply for a lattice light sheet platform that will enable fast and gentle multi-color experiments over extended periods of time. The lattice light sheet technology for the first time allows for long-term multicolor imaging of fast biological processes with subcellular resolution. This technology will thus be the basis for the investigation of many highly relevant biological processes. The platform will be set up in the Light Microscopy Facility (LMF) of the Center for Molecular and Cellular Bioengineering (CMCB) at the TU Dresden, thus securing open access and best-possible support for all users.