| |  Online-Ressource |
|---|
| Verfasst von: | Eismann, Björn [VerfasserIn]  |
|---|
| | Krieger, Teresa G. [VerfasserIn] |
|---|
| | Beneke, Jürgen [VerfasserIn] |
|---|
| | Bulkescher, Ruben [VerfasserIn] |
|---|
| | Adam, Lukas [VerfasserIn] |
|---|
| | Erfle, Holger [VerfasserIn] |
|---|
| | Herrmann, Carl [VerfasserIn] |
|---|
| | Eils, Roland [VerfasserIn] |
|---|
| | Conrad, Christian [VerfasserIn] |
|---|
| Titel: | Automated 3D light-sheet screening with high spatiotemporal resolution reveals mitotic phenotypes |
|---|
| Verf.angabe: | Björn Eismann, Teresa G. Krieger, Jürgen Beneke, Ruben Bulkescher, Lukas Adam, Holger Erfle, Carl Herrmann, Roland Eils and Christian Conrad |
|---|
| E-Jahr: | 2020 |
|---|
| Jahr: | 1 June 2020 |
|---|
| Umfang: | 11 S. |
|---|
| Fussnoten: | Gesehen am 25.08.2020 |
|---|
| Titel Quelle: | Enthalten in: Journal of cell science |
|---|
| Ort Quelle: | Cambridge : Company of Biologists Limited, 1853 |
|---|
| Jahr Quelle: | 2020 |
|---|
| Band/Heft Quelle: | 133(2020,11) Artikel-Nummer jcs245043, 11 Seiten |
|---|
| ISSN Quelle: | 1477-9137 |
|---|
| Abstract: | 3D cell cultures enable the in vitro study of dynamic biological processes such as the cell cycle, but their use in high-throughput screens remains impractical with conventional fluorescent microscopy. Here, we present a screening workflow for the automated evaluation of mitotic phenotypes in 3D cell cultures by light-sheet microscopy. After sample preparation by a liquid handling robot, cell spheroids are imaged for 24 h in toto with a dual-view inverted selective plane illumination microscope (diSPIM) with a much improved signal-to-noise ratio, higher imaging speed, isotropic resolution and reduced light exposure compared to a spinning disc confocal microscope. A dedicated high-content image processing pipeline implements convolutional neural network-based phenotype classification. We illustrate the potential of our approach using siRNA knockdown and epigenetic modification of 28 mitotic target genes for assessing their phenotypic role in mitosis. By rendering light-sheet microscopy operational for high-throughput screening applications, this workflow enables target gene characterization or drug candidate evaluation in tissue-like 3D cell culture models. |
|---|
| DOI: | doi:10.1242/jcs.245043 |
|---|
| URL: | Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.
Volltext: https://doi.org/10.1242/jcs.245043 |
|---|
| | DOI: https://doi.org/10.1242/jcs.245043 |
|---|
| Datenträger: | Online-Ressource |
|---|
| Sprache: | eng |
|---|
| Sach-SW: | Cell cycle |
|---|
| | cells |
|---|
| | convolutional neural-network |
|---|
| | dna |
|---|
| | fluorescence microscopy |
|---|
| | High-content screening |
|---|
| | human genome |
|---|
| | Light-sheet microscopy |
|---|
| K10plus-PPN: | 172772951X |
|---|
| Verknüpfungen: | → Zeitschrift |
|---|
Automated 3D light-sheet screening with high spatiotemporal resolution reveals mitotic phenotypes / Eismann, Björn [VerfasserIn]; 1 June 2020 (Online-Ressource)
