Fast, volumetric live-cell imaging using high-resolution light-field microscopy
Fast, volumetric live-cell imaging using high-resolution light-field microscopy
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DOI：
10.1101/439315
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发表时间：
2018-10
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通讯作者：
Haoyu Li;Changliang Guo;Deborah M Kim-Holzapfel;Weiyi Li;Y. Altshuller;Bryce C. Schroeder;Wenhao Liu-Wenhao-Li
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作者：
Haoyu Li;Changliang Guo;Deborah M Kim-Holzapfel;Weiyi Li;Y. Altshuller;Bryce C. Schroeder;Wenhao Liu-Wenhao-Li
可视化不同的解剖和功能特征，跨越许多空间尺度，具有高时空分辨率，提供了对生物基本原理的见解。光场显微镜（LFM）最近成为一种无需扫描、可扩展的方法，可以实现高速、体积功能脑成像。考虑到这些在组织水平上的应用前景，在另一个极端，这种高度可扩展的方法在观察单细胞标本的结构和动力学方面具有巨大的潜力。然而，目前LFM的挑战仍然是实现亚细胞水平，近衍射限制的3D空间分辨率。在这里，我们报告了用于活细胞成像的高分辨率LFM (HR-LFM)，在所有三个维度上的分辨率为300-700 nm，成像深度为几微米，体积采集时间为毫秒。我们通过成像各种细胞动力学和结构以及跟踪单个粒子来演示该技术。该方法可能会使LFM成为在多个时空水平上理解生物系统的特别有用的工具。
Visualizing diverse anatomical and functional traits that span many spatial scales with high spatio-temporal resolution provides insights into the fundamentals of living organisms. Light-field microscopy (LFM) has recently emerged as a scanning-free, scalable method that allows for high-speed, volumetric functional brain imaging. Given those promising applications at the tissue level, at its other extreme, this highly-scalable approach holds great potential for observing structures and dynamics in single-cell specimens. However, the challenge remains for current LFM to achieve subcellular level, near-diffraction-limited 3D spatial resolution. Here, we report high-resolution LFM (HR-LFM) for live-cell imaging with a resolution of 300-700 nm in all three dimensions, an imaging depth of several micrometers, and a volume acquisition time of milliseconds. We demonstrate the technique by imaging various cellular dynamics and structures and tracking single particles. The method may advance LFM as a particularly useful tool for understanding biological systems at multiple spatio-temporal levels.