Status: Bibliographieeintrag
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| |  Online-Ressource |
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| Verfasst von: | Gilbert, Daniel [VerfasserIn]  |
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| | Boutros, Michael [VerfasserIn] |
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| Titel: | A protocol for a high-throughput multiplex cell viability assay |
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| Verf.angabe: | Daniel F. Gilbert, Michael Boutros |
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| E-Jahr: | 2016 |
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| Jahr: | 01 September 2016 |
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| Umfang: | 10 S. |
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| Fussnoten: | Gesehen am 10.06.2020 |
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| Titel Quelle: | Enthalten in: High-throughput RNAi screening |
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| Ort Quelle: | New York, NY : Humana Press, 2016 |
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| Jahr Quelle: | 2016 |
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| Band/Heft Quelle: | (2016), Seite 75-84 |
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| ISBN Quelle: | 978-1-4939-6337-9 |
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| Abstract: | High-throughput cell viability assays are broadly used in RNAi and small molecule screening experiments to identify compounds that selectively kill cancer cells or as counter screens to exclude the compounds that have a generic effect on cell growth. While there are several assaying techniques available, cellular fitness is often assessed on the basis of one single and often rather indirect physiological indicator. This can lead to inconsistencies and poor correspondence between cell viability screening experiments, conducted under comparable conditions but with different viability indicators. Multiplexing, i.e., the combination of different individual assaying techniques in one experiment and subsequent comparative analysis of multiparametric data can decrease inter-assay variability and increase dataset concordance. Here, we describe a protocol for a multiplexing approach for high-throughput cell viability screening to address the issues encountered in the classical strategy using a single fitness indicator described above. The method combines a biochemical, luminescence-based approach and two fluorescence-based assay types. The biochemical method assesses cellular fitness by quantifying intracellular ATP concentration. Calcein labeling reflects cell fitness through membrane integrity and indirect measurement of ATP-dependent enzymatic esterase activity. Hoechst DNA stain correlates cell fitness with cellular DNA content. The presented multiplexing approach is suitable for low, medium and high-throughput screening and has the potential to decrease inter-assay variability and increase dataset concordance as well as reproducibility of experimental results. |
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| DOI: | doi:10.1007/978-1-4939-6337-9_6 |
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| URL: | Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.
Volltext: https://doi.org/10.1007/978-1-4939-6337-9_6 |
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| | Volltext: https://experiments.springernature.com/articles/10.1007/978-1-4939-6337-9_6 |
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| | DOI: https://doi.org/10.1007/978-1-4939-6337-9_6 |
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| Datenträger: | Online-Ressource |
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| Sprache: | eng |
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| Sach-SW: | Adenosine Triphosphate |
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| | Cell fitness |
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| | Cell Survival |
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| | Cell viability |
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| | Cell-based assays |
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| | Cytological Techniques |
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| | Drug discovery |
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| | HEK293 Cells |
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| | High-throughput screening |
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| | High-Throughput Screening Assays |
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| | Humans |
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| | In vitro toxicity screening |
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| | Multiplexing |
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| | RNA, Small Interfering |
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| | RNAi screening |
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| | Target validation |
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| | Transfection |
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| K10plus-PPN: | 1700338412 |
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| Verknüpfungen: | → Sammelwerk |
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¬A¬ protocol for a high-throughput multiplex cell viability assay / Gilbert, Daniel [VerfasserIn]; 01 September 2016 (Online-Ressource)
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