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Techniques to Enable 1536-Well Phenotypic Screening

  • Sinéad Knight
  • Helen Plant
  • Lisa McWilliams
  • Mark Wigglesworth
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1787)

Abstract

Adaptation of phenotypic cell assays to 1536-well format brings major challenges in liquid handling for high-content assays requiring washing steps and coating of plates. In addition, problematic edge effects and reduced assay quality are frequently encountered. In this chapter, we describe the novel application of a centrifugal plate washer to facilitate miniaturization of 1536-well cell assays and a combination of techniques to reduce edge effects, all of which improved throughput and data quality. Cell assays currently limited in throughput because of cost and complex protocols may be enabled by the techniques presented in this chapter.

Key words

Miniaturization 1536-well High-throughput Cell assays Phenotypic screening 

References

  1. 1.
    Vincent F, Loria P, Pregel M, Stanton R, Kitching L, Nocka K et al (2015) Developing predictive assays: the phenotypic screening “rule of 3”. Sci Transl Med 7:293ps15CrossRefGoogle Scholar
  2. 2.
    Fuller JA, Shaw GC, Bonnet-Wersinger D, Hansen BS, Berlincke CA, Inglese J et al (2014) A high content screening approach to identify molecules neuroprotective for photoreceptor cells. Adv Exp Med Biol 801:773–781CrossRefGoogle Scholar
  3. 3.
    Li J, Crowley ST, Duskey J, Khargharia S, Wu M, Rice KG (2015) Miniaturization of gene transfection assays in 384- and 1536-well microplates. Anal Biochem 470:14–21CrossRefGoogle Scholar
  4. 4.
    Luense S, Denner P, Fernández-Montalván A, Hartung I, Husemann M, Stresemann C et al (2015) Quantification of histone H3 Lys27 trimethylation (H3K27me3) by high-throughput microscopy enables cellular large-scale screening for small-molecule EZH2 inhibitors. J Biomol Screen 20:190–201CrossRefGoogle Scholar
  5. 5.
    Knight S, Plant H, McWilliams L, Murray D, Dixon-Steele R, Varghese A et al (2016) Enabling 1536-well high-throughput cell-based screening through the application of novel centrifugal plate washing. SLAS Discov 22:732–742PubMedGoogle Scholar
  6. 6.
    Zhang JH, Chung TD, Oldenburg KR (1999) A simple statistical parameter for use in evaluation and validation of high throughput screening assays. J Biomol Screen 4:67–73CrossRefGoogle Scholar
  7. 7.
  8. 8.
    http://cn.agilent.com/cs/library/technicaloverviews/public/5991-8020EN.pdf. A quality control procedure for the Agilent Bravo platform

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Sinéad Knight
    • 1
  • Helen Plant
    • 2
  • Lisa McWilliams
    • 1
  • Mark Wigglesworth
    • 2
  1. 1.Discovery Sciences, IMED Biotech UnitAstraZenecaCambridgeUK
  2. 2.Discovery Sciences, IMED Biotech UnitAstraZenecaMacclesfieldUK

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