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Recombinant Differential Anchorage Probes that Tower over the Spatial Dimension of Intracellular Signals for High Content Screening and Analysis

Authors
  • Schembri, Laura
  • Zanese, Marion
  • Depierre-Plinet, Gaelle
  • Petit, Muriel
  • Elkaoukabi-Chaibi, Assia
  • Tauzin, Loic
  • Cristina Florean
  • Lartigue, Lydia
  • Medina, Chantal
  • Rey, Christophe
  • Belloc, Francis
  • Reiffers, Josy
  • Ichas, FrançOis
  • De Giorgi, Francesca
Type
Published Article
Journal
Analytical Chemistry
Publisher
American Chemical Society
Publication Date
Dec 24, 2009
Volume
81
Issue
23
Pages
9598–9598
Identifiers
DOI: 10.1021/ac9015227
Source
LBMCC
Keywords
License
White

Abstract

Recombinant fluorescent probes allow the detection of molecular events inside living cells. Many of them exploit the intracellular space to provide positional signals and, thus, require detection by single cell imaging. We describe here a novel strategy based on probes capable of encoding the spatial dimension of intracellular signals into "all-or-none" fluorescence intensity changes (differential anchorage probes, DAPs). The resulting signals can be acquired in single cells at high throughput by automated flow cytometry, (i) bypassing image acquisition and analysis, (ii) providing a direct quantitative readout, and (iii) allowing the exploration of large experimental series. We illustrate our purpose with DAPs for Bax and the effector caspases 3 and 7, which are keys players in apoptotic cell death, and show applications in basic research, high content multiplexed library screening, compound characterization, and drug profiling.

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