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

• Type of Material: Article
• Authors: Zhi Ling, Emory University; Keyi Han, Emory University; Liu Wenhau, Emory University; Xuanwen Hua, Emory University; Shu Jia, Emory University
• Language: English
• Date: 2023-07-25
• Publisher: Optica Publishing
• Publication Version: Final Published Version
• Copyright Statement: © 2023 Optica Publishing Group under the terms of the Optica Open Access Publishing Agreement
• License: Creative Commons Attribution-NonCommercial 4.0 International
• Final Published Version (URL): https://doi.org/10.1364/BOE.495506
• Title of Journal or Parent Work: Biomedical Optics
• Volume: 14
• Issue: 8
• Start Page: 4237
• End Page: 4245
• Grant/Funding Information: National Science Foundation10.13039/100000001 (DBI2145235, EFMA1830941); National Institutes of Health10.13039/100000002 (R35GM124846).
• Keywords: microscopy; sub-cellular structures; volumetric live-cell imaging; HeLa Cells; Fourier light-field microscopy
• Research Categories: Health Sciences, Opthamology; Biology, Cell

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.