Isotropic imaging across spatial scales with axially swept light-sheet microscopy.
Isotropic imaging across spatial scales with axially swept light-sheet microscopy.
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DOI：
10.1038/s41596-022-00706-6
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发表时间：
2022-09
期刊：
影响因子：
14.8
通讯作者：
Fiolka, Reto
中科院分区：
文献类型：
--
作者：
Dean, Kevin M.;Chakraborty, Tonmoy;Daetwyler, Stephan;Lin, Jinlong;Garrelts, Gerard;M'Saad, Ons;Mekbib, Hannahmariam T.;Voigt, Fabian F.;Schaettin, Martina;Stoeckli, Esther T.;Helmchen, Fritjof;Bewersdorf, Joerg;Fiolka, Reto
光片荧光显微镜(LSFM)是一种发展迅速的技术，由于其高时空分辨率和温和的无光毒性光照，在生命科学中得到了极大的普及。在该协议中，我们为多功能LSFM的组装和操作提供了详细的指导，称为轴向扫描光片显微镜(ASLM)，它提供了视场、光学分辨率和光学切片的无与伦比的组合。为了使ASLM大众化，我们概述了它的工作原理、在生物成像中的应用，以及对其光学系统的组装、对准和控制的实用提示。此外，我们还提供了几种ASLM变体的详细部件列表和原理图，这些部件列表和原理图结合在一起，可以解析化学膨胀样品中的分子细节、活细胞中的亚细胞组织，或化学清除后的完整生物体的解剖组成。我们还提供仪器控制软件，并讨论用户如何调整成像参数以适应不同的样本类型。因此，该协议不仅将作为采用ASLM的入门和高级用户的指南，而且将作为任何对部署定制成像技术感兴趣的个人的有用资源。我们预计，建立ASLM的时间将需要大约1-2个月，这取决于仪器制造商的经验和仪器的版本。本协议描述了轴向扫描薄片显微镜的工作原理和实现方法，它可以在器官尺度上对亚细胞特征进行3D成像。讨论了如何优化其设计，并为其施工和线形提供了详细的指导。
Light-Sheet Fluorescence Microscopy (LSFM) is a rapidly growing technique that has gained tremendous popularity in the life sciences owing to its high-spatiotemporal resolution and gentle, non-phototoxic illumination. In this protocol, we provide detailed directions for the assembly and operation of a versatile LSFM variant, referred to as Axially Swept Light-Sheet Microscopy (ASLM), that delivers an unparalleled combination of field of view, optical resolution, and optical sectioning. To democratize ASLM, we provide an overview of its working principle, applications to biological imaging, as well as pragmatic tips for the assembly, alignment, and control of its optical systems. Furthermore, we provide detailed part lists and schematics for several variants of ASLM, that together, can resolve molecular detail in chemically expanded samples, subcellular organization in living cells, or the anatomical composition of chemically cleared intact organisms. We also provide software for instrument control and discuss how users can tune imaging parameters to accommodate diverse sample types. Thus, this protocol will serve not only as a guide for both introductory and advanced users adopting ASLM, but as a useful resource for any individual interested in deploying custom imaging technology. We expect that the time to build an ASLM will take about 1–2 months, depending on the experience of the instrument builder and the version of the instrument. This protocol describes the working principle and implementations of Axially Swept Light-Sheet Microscopy, which can image subcellular features at organ scales in 3D. It discusses how to optimize its design, and provides a detailed guide for its construction and alignment.
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DOI：
10.1038/s41377-020-00401-9
发表时间：
2020
期刊：
Light, science & applications
影响因子：
--
作者：
Chakraborty T;Chen B;Daetwyler S;Chang BJ;Vanderpoorten O;Sapoznik E;Kaminski CF;Knowles TPJ;Dean KM;Fiolka R
通讯作者：
Fiolka R
影响因子：
44.1
作者：
Jing D;Zhang S;Luo W;Gao X;Men Y;Ma C;Liu X;Yi Y;Bugde A;Zhou BO;Zhao Z;Yuan Q;Feng JQ;Gao L;Ge WP;Zhao H
通讯作者：
Zhao H
影响因子：
3.4
作者：
Dean, Kevin M.;Roudot, Philippe;Welf, Erik S.;Danuser, Gaudenz;Fiolka, Reto
通讯作者：
Fiolka, Reto
影响因子：
48
作者：
Chhetri, Raghav K.;Amat, Fernando;Kellerbeams, Philipp J.
通讯作者：
Kellerbeams, Philipp J.
影响因子：
3.8
作者：
Chang, Bo-Jui;Dean, Kevin M.;Fiolka, Reto
通讯作者：
Fiolka, Reto