Delyan P. Ivanov
Spheroid arrays for high-throughput single-cell analysis of spatial patterns and biomarker expression in 3D
Ivanov, Delyan P.; Grabowska, Anna M.
Abstract
We describe and share a device, methodology and image analysis algorithms, which allow up to 66 spheroids to be arranged into a gel-based array directly from a culture plate for downstream processing and analysis. Compared to processing individual samples, the technique uses 11-fold less reagents, saves time and enables automated imaging. To illustrate the power of the technology, we showcase applications of the methodology for investigating 3D spheroid morphology and marker expression and for in vitro safety and efficacy screens. Firstly, spheroid arrays of 11 cell-lines were rapidly assessed for differences in spheroid morphology. Secondly, highly-positive (SOX-2), moderately-positive (Ki-67) and weakly-positive (?III-tubulin) protein targets were detected and quantified. Third, the arrays enabled screening of ten media compositions for inducing differentiation in human neurospheres. Lastly, the application of spheroid microarrays for spheroid-based drug-screens was demonstrated by quantifying the dose-dependent drop in proliferation and increase in differentiation in etoposide-treated neurospheres.
Citation
Ivanov, D. P., & Grabowska, A. M. (2017). Spheroid arrays for high-throughput single-cell analysis of spatial patterns and biomarker expression in 3D. Scientific Reports, 7(1), Article 41160. https://doi.org/10.1038/srep41160
Journal Article Type | Article |
---|---|
Acceptance Date | Dec 14, 2016 |
Online Publication Date | Jan 30, 2017 |
Publication Date | 2017-12 |
Deposit Date | Jan 17, 2017 |
Publicly Available Date | Mar 29, 2024 |
Journal | Scientific Reports |
Electronic ISSN | 2045-2322 |
Publisher | Nature Publishing Group |
Peer Reviewed | Peer Reviewed |
Volume | 7 |
Issue | 1 |
Article Number | 41160 |
DOI | https://doi.org/10.1038/srep41160 |
Public URL | https://nottingham-repository.worktribe.com/output/838657 |
Publisher URL | http://www.nature.com/articles/srep41160 |
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Copyright Statement
Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0
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