Simultaneous Multicolor Multifocal Scanning Microscopy

• Type of Material: Article
• Authors: Kyungduck Yoon, Emory University; Keyi Han, Emory University; Kidan Tadesse, Emory University; Biagio Mandracchia, Emory University; Shu Jia, Emory University
• Language: English
• Date: 2023-07-24
• Publisher: American Chemical Society
• Publication Version: Final Published Version
• Copyright Statement: © 2023 The Authors. Published by American Chemical Society
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): https://doi.org/10.1021%2Facsphotonics.3c00205
• Title of Journal or Parent Work: American Chemical Society Photonics
• Volume: 10
• Issue: 9
• Start Page: 3035
• End Page: 3041
• Grant/Funding Information: This work is supported by the Parker H. Petit Institute for Bioengineering and Biosciences of Georgia Institute of Technology, NIH grant R35GM124846, and NSF grants EFMA1830941 and DBI2145235.
• Keywords: MSM; super-resolution systems; optical sectioning; cell biology; image-scanning microscopy; super-resolution imaging
• Research Categories: Biology, Cell

Abstract

Super-resolution fluorescence microscopy has revolutionized cell biology over the past decade, enabling the visualization of subcellular complexity with unparalleled clarity and detail. However, the rapid development of image-scanning-based super-resolution systems still restrains convenient access to commonly used instruments such as epi-fluorescence microscopes. Here, we present multifocal scanning microscopy (MSM) for super-resolution imaging with simultaneous multicolor acquisition and minimal instrumental complexity. MSM implements a stationary, interposed multifocal multicolor excitation by exploiting the motion of the specimens, realizing super-resolution microscopy through a general epi-fluorescence platform without compromising the image-scanning mechanism or inducing complex instrument alignment. The system is demonstrated with various phantom and biological specimens, and the results present effective resolution doubling, optical sectioning, and contrast enhancement. We anticipate MSM, as a highly accessible and compatible super-resolution technique, to offer a promising methodological pathway for broad cell biological discoveries.