| Download | - View final version: An automated screening platform for improving the responsiveness of genetically encoded Ca²⁺ biosensors in mammalian cells (PDF, 2.9 MiB)
- View supplementary information: An automated screening platform for improving the responsiveness of genetically encoded Ca²⁺ biosensors in mammalian cells (PDF, 347 KiB)
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| DOI | Resolve DOI: https://doi.org/10.1039/D4SD00138A |
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| Author | Search for: Zhao, Yufeng1ORCID identifier: https://orcid.org/0000-0002-1788-7969; Search for: Shen, YiORCID identifier: https://orcid.org/0000-0003-4117-0279; Search for: Veres, Teodor1ORCID identifier: https://orcid.org/0000-0002-9594-8042; Search for: Campbell, Robert E. |
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| Affiliation | - National Research Council of Canada. Medical Devices
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| Funder | Search for: Natural Sciences and Engineering Research Council of Canada; Search for: Japan Society for the Promotion of Science; Search for: Canadian Institutes of Health Research |
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| Format | Text, Article |
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| Abstract | Genetically-encoded, fluorescent protein (FP)-based biosensors are powerful tools for imaging dynamic cellular activities. Directed evolution is a highly effective method for developing enhanced versions of FP-based biosensors, but the screening process is laborious and time-consuming. Mammalian cell-based screening with electrical stimulation methods has been successful in accurately selecting variants of biosensors for imaging neuronal activities. We introduce an automated mammalian cell screening platform utilizing a fluorescence microscope and a liquid dispenser to enable the screening of biosensor responsiveness to chemical stimulation. We demonstrated the effectiveness of this platform in improving the response of a red fluorescent biosensor for Ca²⁺, K-GECO, for detection of histamine-induced changes in Ca²⁺ concentration. This method should be applicable to any FP-based biosensor that responds to pharmacological treatment or other exogenous chemical stimulation, simplifying efforts to develop biosensors tailored for specific applications in diverse biological contexts. |
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| Publication date | 2024-07-18 |
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| Publisher | Royal Society of Chemistry |
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| Licence | |
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| In | |
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| Language | English |
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| Peer reviewed | Yes |
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| Export citation | Export as RIS |
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| Report a correction | Report a correction (opens in a new tab) |
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| Record identifier | b3cfd84f-4c34-49a3-b0ab-72ec9f76cdf9 |
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| Record created | 2025-06-12 |
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| Record modified | 2025-06-12 |
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