Navigation überspringen
Universitätsbibliothek Heidelberg
Status: Bibliographieeintrag
Standort: ---
Exemplare: ---
heiBIB
 Online-Ressource
Verfasst von:Gilbert, Daniel [VerfasserIn]   i
 Boutros, Michael [VerfasserIn]   i
Titel:A protocol for a high-throughput multiplex cell viability assay
Verf.angabe:Daniel F. Gilbert, Michael Boutros
E-Jahr:2016
Jahr:01 September 2016
Umfang:10 S.
Fussnoten:Gesehen am 10.06.2020
Titel Quelle:Enthalten in: High-throughput RNAi screening
Ort Quelle:New York, NY : Humana Press, 2016
Jahr Quelle:2016
Band/Heft Quelle:(2016), Seite 75-84
ISBN Quelle:978-1-4939-6337-9
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.
DOI:doi:10.1007/978-1-4939-6337-9_6
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
 Volltext: https://experiments.springernature.com/articles/10.1007/978-1-4939-6337-9_6
 DOI: https://doi.org/10.1007/978-1-4939-6337-9_6
Datenträger:Online-Ressource
Sprache:eng
Sach-SW:Adenosine Triphosphate
 Cell fitness
 Cell Survival
 Cell viability
 Cell-based assays
 Cytological Techniques
 Drug discovery
 HEK293 Cells
 High-throughput screening
 High-Throughput Screening Assays
 Humans
 In vitro toxicity screening
 Multiplexing
 RNA, Small Interfering
 RNAi screening
 Target validation
 Transfection
K10plus-PPN:1700338412
Verknüpfungen:→ Sammelwerk

Permanenter Link auf diesen Titel (bookmarkfähig):  https://katalog.ub.uni-heidelberg.de/titel/68586101   QR-Code
zum Seitenanfang