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

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• 作者：Laura Schembri ; Marion Zanese ; Gaelle Depierre-Plinet ; Muriel Petit ; Assia Elkaoukabi-Chaibi ; Loic Tauzin ; Cristina Florean ; Lydia Lartigue ; Chantal Medina ; Christophe Rey ; Francis Belloc ; Josy Reiffe
• 刊名：Analytical Chemistry
• 出版年：2009
• 出版时间：December 1, 2009
• 年：2009
• 卷：81
• 期：23
• 页码：9590-9598
• 全文大小：484K
• 年卷期：v.81,no.23(December 1, 2009)
• ISSN：1520-6882

文摘

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.