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使用傅里叶光场显微镜进行体积活细胞自发荧光成像。

Volumetric live-cell autofluorescence imaging using Fourier light-field microscopy.

作者信息

Ling Zhi, Han Keyi, Liu Wenhao, Hua Xuanwen, Jia Shu

机构信息

The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332, USA.

Parker H. Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, GA 30332, USA.

出版信息

Biomed Opt Express. 2023 Jul 25;14(8):4237-4245. doi: 10.1364/BOE.495506. eCollection 2023 Aug 1.

Abstract

This study introduces a rapid, volumetric live-cell imaging technique for visualizing autofluorescent sub-cellular structures and their dynamics by employing high-resolution Fourier light-field microscopy. We demonstrated this method by capturing lysosomal autofluorescence in fibroblasts and HeLa cells. Additionally, we conducted multicolor imaging to simultaneously observe lysosomal autofluorescence and fluorescently-labeled organelles such as lysosomes and mitochondria. We further analyzed the data to quantify the interactions between lysosomes and mitochondria. This research lays the foundation for future exploration of native cellular states and functions in three-dimensional environments, effectively reducing photodamage and eliminating the necessity for exogenous labels.

摘要

本研究介绍了一种快速的体视活细胞成像技术,该技术通过使用高分辨率傅里叶光场显微镜来可视化自发荧光亚细胞结构及其动态变化。我们通过捕捉成纤维细胞和HeLa细胞中的溶酶体自发荧光来证明了该方法。此外,我们进行了多色成像,以同时观察溶酶体自发荧光以及荧光标记的细胞器,如溶酶体和线粒体。我们进一步分析数据以量化溶酶体与线粒体之间的相互作用。这项研究为未来在三维环境中探索天然细胞状态和功能奠定了基础,有效减少了光损伤并消除了对外源标记的需求。

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