Confocal-based method for quantification of diffusion kinetics in microwell plates and its application for identifying a rapid mixing method for high-content/throughput screening

Ok Ryul Song, Tae Hee Kim, Xavier Perrodon, Changbok Lee, Hee Kyoung Jeon, Zahir Seghiri, Ho Jeong Kwon, Jonathan Cechetto, Thierry Christophe

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Rapid mixing in microplates is still an underappreciated challenge in screening assay development, particularly with the use of noncontact nanoliter liquid handlers. In high-content/throughput screening (HC/TS), fast and efficient mixing between compounds and cell culture medium is even more critical as biological kinetics dictates speed of mixing, usually within a few minutes. Moreover, mixing in HC/TS should be gentle enough, to avoid any negative disruption in. cell layer. Here the authors introduce a method to accurately quantify drop diffusion into a microplate well, independently of buffer, liquid han-dler, or dispensing protocol. This method was used to determine the effect of various mixing methods on the diffusion of a nanoliter drop of pure DMSO in aqueous buffer in 384-well plates. Rapid plate shaking and additional buffer addition were shown to be the most efficient and effective mixing methods for HC/TS. However, efficient mixing by plate shaking is lim-ited by assay volume. Bulk addition shows fast and efficient mixing, without negative effects on cells. Moreover, this simple, fast, and inexpensive method can be easily adapted on any platform.

Original languageEnglish
Pages (from-to)138-147
Number of pages10
JournalJournal of Biomolecular Screening
Volume15
Issue number2
DOIs
Publication statusPublished - 2010

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biotechnology
  • Biochemistry
  • Molecular Medicine
  • Pharmacology
  • Drug Discovery

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