期刊
SCIENTIFIC REPORTS
卷 10, 期 1, 页码 -出版社
NATURE PORTFOLIO
DOI: 10.1038/s41598-020-58563-w
关键词
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资金
- Helmholtz Association program BioInterfaces in Technology and Medicine - BIFTM
- European Research Council [294354-ManISteC, ERC-2018-SyG-810172]
- European Union's Horizon 2020 research and innovation program 4Dheart [722427]
- Research Center for Molecular Medicine (HRCMM) Career Development Fellowship (CDF)
- MD/PhD program of the Medical Faculty Heidelberg
- Deutsche Herzstiftung e.V. [S/02/17]
- Add-On Fellowship for Interdisciplinary Science of the Joachim Herz Stiftung
- Deutsches Zentrum fur Herz-Kreislauf-Forschung (DZHK)
- Marie Curie Actions (MSCA) [722427] Funding Source: Marie Curie Actions (MSCA)
Accurate quantification of heartbeats in fish models is an important readout to study cardiovascular biology, disease states and pharmacology. However, dependence on anaesthesia, laborious sample orientation or requirement for fluorescent reporters have hampered the use of high-throughput heartbeat analysis. To overcome these limitations, we established an efficient screening assay employing automated label-free heart rate determination of randomly oriented, non-anesthetized medaka (Oryzias latipes) and zebrafish (Danio rerio) embryos in microtiter plates. Automatically acquired bright-field data feeds into an easy-to-use HeartBeat software with graphical user interface for automated quantification of heart rate and rhythm. Sensitivity of the assay was demonstrated by profiling heart rates during entire embryonic development. Our analysis revealed rapid adaption of heart rates to temperature changes, which has implications for standardization of experimental layout. The assay allows scoring of multiple embryos per well enabling a throughput of >500 embryos per 96-well plate. In a proof of principle screen for compound testing, we captured concentration-dependent effects of nifedipine and terfenadine over time. Our novel assay permits large-scale applications ranging from phenotypic screening, interrogation of gene functions to cardiovascular drug development.
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