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Published (Version of record)CC BY V4.0, Open Access
Published (Version of record)CC BY V4.0, Open Access
Journal article
Circa-SCOPE: high-throughput live single-cell imaging method for analysis of circadian clock resetting
Nature communications, Vol.12(1), 5903
08/Oct/2021
Abstract
Circadian clocks are self-sustained and cell-autonomous oscillators. They respond to various extracellular cues depending on the time-of-day and the signal intensity. Phase Transition Curves (PTCs) are instrumental in uncovering the full repertoire of responses to a given signal. However, the current methodologies for reconstructing PTCs are low-throughput, laborious, and resource- and time-consuming. We report here the development of an efficient and high throughput assay, dubbed Circadian Single-Cell Oscillators PTC Extraction (Circa-SCOPE) for generating high-resolution PTCs. This methodology relies on continuous monitoring of single-cell oscillations to reconstruct a full PTC from a single culture, upon a one-time intervention. Using Circa-SCOPE, we characterize the effects of various pharmacological and blood-borne resetting cues, at high temporal resolution and a wide concentration range. Thus, Circa-SCOPE is a powerful tool for comprehensive analysis and screening for circadian clocks’ resetting cues, and can be valuable for basic as well as translational research.
Details
- Title
- Circa-SCOPE; high-throughput live single-cell imaging method for analysis of circadian clock resetting
- Creators
- Gal Manella - 972WIS_INST___112Dan Aizik - 972WIS_INST___112Rona Aviram - 972WIS_INST___112Marina Golik - 972WIS_INST___112Gad Asher - 972WIS_INST___112
- Resource Type
- Journal article
- Publication Details
- Nature communications, Vol.12(1), 5903; 08/Oct/2021
- Number of pages
- 14
- Language
- English
- DOI
- https://doi.org/10.1038/s41467-021-26210-1
- Grant note
- We are grateful to all the members of the Asher lab for their comments on the paper. We wish to thank Uriel Rosen (WIS IT unit) and Jonathan Sobel for their assistance in establishing the CellProfiler pipeline on the institutional cluster. G.A. is supported by a grant from the European Research Council (ERC-2017 CIRCOMMUNICATION 770869), Abisch Frenkel Foundation for the Promotion of Life Sciences, Adelis Foundation, Susan and Michael Stern. Author contributions : Conceptualization: G.M. and G.A.; Investigation: G.M., D.A., R.A. and M.G.; Visualization: G.M.; Data curation: G.M.; Software: G.M.; Funding: G.A.; Writing—review & editing: G.M. and G.A.
- Record Identifier
- 993320871503596
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