Circa-SCOPE: high-throughput live single-cell imaging method for analysis of circadian clock resetting

Gal Manella, Dan Aizik, Rona Aviram, Marina Golik, Gad Asher

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

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.

Original languageEnglish
Article number5903
JournalNature Communications
Volume12
Issue number1
DOIs
StatePublished - 8 Oct 2021
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2021, The Author(s).

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