Multicolor multifocal 3D microscopy using in-situ optimization of a spatial light modulator.

TitleMulticolor multifocal 3D microscopy using in-situ optimization of a spatial light modulator.
Publication TypeJournal Article
Year of Publication2022
AuthorsM Amin, J, Zhao, T, Yang, H, Shaevitz, JW
JournalSci Rep
Date Published2022 Sep 29
KeywordsEscherichia coli, Escherichia coli Infections, Fluorescent Dyes, Humans, Lighting, Microscopy

<p>Multifocal microscopy enables high-speed three-dimensional (3D) volume imaging by using a multifocal grating in the emission path. This grating is typically designed to afford a uniform illumination of multifocal subimages for a single emission wavelength. Using the same grating for multicolor imaging results in non-uniform subimage intensities in emission wavelengths for which the grating is not designed. This has restricted multifocal microscopy applications for samples having multicolored fluorophores. In this paper, we present a multicolor multifocal microscope implementation which uses a Spatial Light Modulator (SLM) as a single multifocal grating to realize near-uniform multifocal subimage intensities across multiple wavelength emission bands. Using real-time control of an in-situ-optimized SLM implemented as a multifocal grating, we demonstrate multicolor multifocal 3D imaging over three emission bands by imaging multicolored particles as well as Escherichia coli (E. coli) interacting with human liver cancer cells, at [Formula: see text] multicolor 3D volumes per second acquisition speed. Our multicolor multifocal method is adaptable across SLM hardware, emission wavelength band locations and number of emission bands, making it particularly suited for researchers investigating fast processes occurring across a volume where multiple species are involved.</p>

Alternate JournalSci Rep
PubMed ID36175472
PubMed Central IDPMC9522655
Grant ListDE-SC0019364 / / the U.S. Department of Energy /
DE-SC0019364 / / the U.S. Department of Energy /