Light‐sheet fluorescence microscopy (LSFM) allows volumetric live imaging at high‐speed and with low photo‐toxicity. Various LSFM modalities are commercially available, but their size and cost limit their access by the research community. A new method, termed sub‐voxel‐resolving (SVR) light‐sheet add‐on microscopy (SLAM), is presented to enable fast, resolution‐enhanced light‐sheet fluorescence imaging from a conventional wide‐field microscope. This method contains two components: a miniature add‐on device to regular wide‐field microscopes, which contains a horizontal laser light‐sheet illumination path to confine fluorophore excitation at the vicinity of the focal plane for optical sectioning; an off‐axis scanning strategy and a SVR algorithm that utilizes sub‐voxel spatial shifts to reconstruct the image volume that results in a twofold increase in resolution. SLAM method has been applied to observe the muscle activity change of crawling C. elegans, the heartbeat of developing zebrafish embryo, and the neural anatomy of cleared mouse brains, at high spatiotemporal resolution. It provides an efficient and cost‐effective solution to convert the vast number of in‐service microscopes for fast 3D live imaging with voxel‐super‐resolved capability.

中文翻译：

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通过亚体素分解光片附加显微镜进行高效且经济高效的3D细胞成像。

薄板荧光显微镜（LSFM）允许高速，低光毒性的实时体积成像。各种LSFM模式都可以通过商业途径获得，但是它们的大小和成本限制了研究社区的访问。提出了一种称为亚体素分解（SVR）光学片附加显微镜（SLAM）的新方法，可以通过常规的宽视场显微镜实现快速，增强分辨率的光学片荧光成像。这种方法包括两个部分：一个用于常规宽视野显微镜的微型附加装置，该装置包含一个水平的激光薄片照明路径，以将荧光团激发限制在焦平面附近以进行光学切片；一种离轴扫描策略和一种SVR算法，该算法利用体素下的空间移位来重建图像体积，从而使分辨率提高了两倍。SLAM方法已被应用于观察爬行的肌肉活动变化Ç。线虫，正在发展的斑马鱼胚胎的心跳，以及在高时空分辨率下清晰的老鼠大脑的神经解剖结构。它提供了一种高效且具有成本效益的解决方案，可将大量在用显微镜转换为具有体素超分辨功能的快速3D实时成像。