An integrated platform for high-throughput nanoscopy.

Autor: Barentine AES; Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA.; Department of Biomedical Engineering, Yale University, New Haven, CT, USA., Lin Y; Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA.; Department of Biomedical Engineering, Yale University, New Haven, CT, USA., Courvan EM; Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA.; Department of Molecular Biophysics and Biochemistry, Yale School of Medicine, New Haven, CT, USA., Kidd P; Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA., Liu M; Department of Genetics, Yale School of Medicine, New Haven, CT, USA., Balduf L; Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA.; Department of Computer Science and Mathematics, University of Applied Sciences, Munich, Germany., Phan T; Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA.; Department of Computer Science and Mathematics, University of Applied Sciences, Munich, Germany., Rivera-Molina F; Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA., Grace MR; Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA., Marin Z; Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA.; Department of Biomedical Engineering, Yale University, New Haven, CT, USA.; Auckland Bioengineering Institute at University of Auckland, Auckland, New Zealand., Lessard M; Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA., Rios Chen J; Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA., Wang S; Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA.; Department of Genetics, Yale School of Medicine, New Haven, CT, USA., Neugebauer KM; Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA.; Department of Molecular Biophysics and Biochemistry, Yale School of Medicine, New Haven, CT, USA., Bewersdorf J; Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA. joerg.bewersdorf@yale.edu.; Department of Biomedical Engineering, Yale University, New Haven, CT, USA. joerg.bewersdorf@yale.edu.; Department of Physics, Yale University, New Haven, CT, USA. joerg.bewersdorf@yale.edu.; Nanobiology Institute, Yale University, West Haven, CT, USA. joerg.bewersdorf@yale.edu., Baddeley D; Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA. d.baddeley@auckland.ac.nz.; Auckland Bioengineering Institute at University of Auckland, Auckland, New Zealand. d.baddeley@auckland.ac.nz.; Nanobiology Institute, Yale University, West Haven, CT, USA. d.baddeley@auckland.ac.nz.
Jazyk: angličtina
Zdroj: Nature biotechnology [Nat Biotechnol] 2023 Nov; Vol. 41 (11), pp. 1549-1556. Date of Electronic Publication: 2023 Mar 13.
DOI: 10.1038/s41587-023-01702-1
Abstrakt: Single-molecule localization microscopy enables three-dimensional fluorescence imaging at tens-of-nanometer resolution, but requires many camera frames to reconstruct a super-resolved image. This limits the typical throughput to tens of cells per day. While frame rates can now be increased by over an order of magnitude, the large data volumes become limiting in existing workflows. Here we present an integrated acquisition and analysis platform leveraging microscopy-specific data compression, distributed storage and distributed analysis to enable an acquisition and analysis throughput of 10,000 cells per day. The platform facilitates graphically reconfigurable analyses to be automatically initiated from the microscope during acquisition and remotely executed, and can even feed back and queue new acquisition tasks on the microscope. We demonstrate the utility of this framework by imaging hundreds of cells per well in multi-well sample formats. Our platform, implemented within the PYthon-Microscopy Environment (PYME), is easily configurable to control custom microscopes, and includes a plugin framework for user-defined extensions.
(© 2023. The Author(s), under exclusive licence to Springer Nature America, Inc.)
Databáze: MEDLINE
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