High-Throughput Video Processing of Heart Rate Responses in Multiple Wild-type Embryonic Zebrafish per Imaging Field
| Autor: | Stephanie Padilla, W. Kyle Martin, Wayne E. Cascio, Michael D. Hays, David M. DeMarini, M. Ian Gilmour, Joey S Stevens, Katya M. Prince, Aimen K. Farraj, Rory B. Conolly, Leslie C. Thompson, Alan H. Tennant, Brandi L Martin, Mehdi S. Hazari |
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| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Embryo Nonmammalian Drug Evaluation Preclinical lcsh:Medicine Image processing Biology Article 03 medical and health sciences 0302 clinical medicine Heart Rate Heart rate Image Processing Computer-Assisted Animals lcsh:Science Throughput (business) Zebrafish Cardiotoxicity Multidisciplinary Cardiac cycle lcsh:R Video processing biology.organism_classification Embryonic stem cell High-Throughput Screening Assays 030104 developmental biology Particulate Matter lcsh:Q 030217 neurology & neurosurgery Biomedical engineering |
| Zdroj: | Scientific Reports, Vol 9, Iss 1, Pp 1-14 (2019) Scientific Reports |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/s41598-018-35949-5 |
| Popis: | Heart rate assays in wild-type zebrafish embryos have been limited to analysis of one embryo per video/imaging field. Here we present for the first time a platform for high-throughput derivation of heart rate from multiple zebrafish (Danio rerio) embryos per imaging field, which is capable of quickly processing thousands of videos and ideal for multi-well platforms with multiple fish/well. This approach relies on use of 2-day post fertilization wild-type embryos, and uses only bright-field imaging, circumventing requirement for anesthesia or restraint, costly software/hardware, or fluorescently-labeled animals. Our original scripts (1) locate the heart and record pixel intensity fluctuations generated by each cardiac cycle using a robust image processing routine, and (2) process intensity data to derive heart rate. To demonstrate assay utility, we exposed embryos to the drugs epinephrine and clonidine, which increased or decreased heart rate, respectively. Exposure to organic extracts of air pollution-derived particulate matter, including diesel or biodiesel exhausts, or wood smoke, all complex environmental mixtures, decreased heart rate to varying degrees. Comparison against an established lower-throughput method indicated robust assay fidelity. As all code and executable files are publicly available, this approach may expedite cardiotoxicity screening of compounds as diverse as small molecule drugs and complex chemical mixtures. |
| Databáze: | OpenAIRE |
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