Increasing the field-of-view in oblique plane microscopy via optical tiling.
Autor: | Chen B; Lyda Hill Department of Bioinformatics, UT Southwestern Medical Center, Dallas, Texas, USA., Chang BJ; Lyda Hill Department of Bioinformatics, UT Southwestern Medical Center, Dallas, Texas, USA., Zhou FY; Lyda Hill Department of Bioinformatics, UT Southwestern Medical Center, Dallas, Texas, USA., Daetwyler S; Lyda Hill Department of Bioinformatics, UT Southwestern Medical Center, Dallas, Texas, USA., Sapoznik E; Lyda Hill Department of Bioinformatics, UT Southwestern Medical Center, Dallas, Texas, USA., Nanes BA; Lyda Hill Department of Bioinformatics, UT Southwestern Medical Center, Dallas, Texas, USA.; Department of Dermatology, UT Southwestern Medical Center, Dallas, Texas, USA., Terrazas I; Department of Internal Medicine, Division of Hematology and Oncology, UT Southwestern Medical Center, Dallas, Texas, USA.; Department of Immunology, UT Southwestern Medical Center, Dallas, Texas, USA.; Simmons Comprehensive Cancer Center, UT Southwestern Medical Center, Dallas, Texas, USA., Gihana GM; Lyda Hill Department of Bioinformatics, UT Southwestern Medical Center, Dallas, Texas, USA., Castro LP; Department of Cell Biology, UT Southwestern Medical Center, Dallas, Texas, USA., Chan IS; Department of Internal Medicine, Division of Hematology and Oncology, UT Southwestern Medical Center, Dallas, Texas, USA.; Simmons Comprehensive Cancer Center, UT Southwestern Medical Center, Dallas, Texas, USA.; Department of Molecular Biology, UT Southwestern Medical Center, Dallas, Texas, USA., Conacci-Sorrell M; Department of Cell Biology, UT Southwestern Medical Center, Dallas, Texas, USA.; Harold C. Simmons Comprehensive Cancer Center, UT Southwestern Medical Center, Dallas, USA.; Hamon Center for Regenerative Science and Medicine, University of Texas Southwestern Medical Center, Dallas, Texas, USA., Dean KM; Lyda Hill Department of Bioinformatics, UT Southwestern Medical Center, Dallas, Texas, USA., Millett-Sikking A; Calico Life Sciences LLC, South San Francisco, CA, USA., York AG; Calico Life Sciences LLC, South San Francisco, CA, USA., Fiolka R; Lyda Hill Department of Bioinformatics, UT Southwestern Medical Center, Dallas, Texas, USA. |
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Jazyk: | angličtina |
Zdroj: | Biomedical optics express [Biomed Opt Express] 2022 Oct 04; Vol. 13 (11), pp. 5616-5627. Date of Electronic Publication: 2022 Oct 04 (Print Publication: 2022). |
DOI: | 10.1364/BOE.467969 |
Abstrakt: | Fast volumetric imaging of large fluorescent samples with high-resolution is required for many biological applications. Oblique plane microscopy (OPM) provides high spatiotemporal resolution, but the field of view is typically limited by its optical train and the pixel number of the camera. Mechanically scanning the sample or decreasing the overall magnification of the imaging system can partially address this challenge, albeit by reducing the volumetric imaging speed or spatial resolution, respectively. Here, we introduce a novel dual-axis scan unit for OPM that facilitates rapid and high-resolution volumetric imaging throughout a volume of 800 × 500 × 200 microns. This enables us to perform volumetric imaging of cell monolayers, spheroids and zebrafish embryos with subcellular resolution. Furthermore, we combined this microscope with a multi-perspective projection imaging technique that increases the volumetric interrogation rate to more than 10 Hz. This allows us to rapidly probe a large field of view in a dimensionality reduced format, identify features of interest, and volumetrically image these regions with high spatiotemporal resolution. Competing Interests: The authors declare no conflicts of interest. (© 2022 Optica Publishing Group under the terms of the Optica Open Access Publishing Agreement.) |
Databáze: | MEDLINE |
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