Molecular-, Cellular- and Pharmacobiology Section, Institute of Pharmaceutical Biology, University of Bonn, Bonn, Germany.
Nat Protoc. 2011 Oct 20;6(11):1748-60. doi: 10.1038/nprot.2011.386.
Label-free dynamic mass redistribution (DMR) is a cutting-edge assay technology that enables real-time detection of integrated cellular responses in living cells. It relies on detection of refractive index alterations on biosensor-coated microplates that originate from stimulus-induced changes in the total biomass proximal to the sensor surface. Here we describe a detailed protocol to apply DMR technology to frame functional behavior of G protein-coupled receptors that are traditionally examined with end point assays on the basis of detection of individual second messengers, such as cAMP, Ca(2+) or inositol phosphates. The method can be readily adapted across diverse cellular backgrounds (adherent or suspension), including primary human cells. Real-time recordings can be performed in 384-well microtiter plates and be completed in 2 h, or they can be extended to several hours depending on the biological question to be addressed. The entire procedure, including cell harvesting and DMR detection, takes 1-2 d.
无标记动态质量重分布（DMR）是一种前沿的检测技术，可实时检测活细胞中整合的细胞反应。它依赖于检测涂有生物传感器的微板上的折射率变化，这些变化源于传感器表面附近总生物量因刺激而发生的变化。在这里，我们描述了一个详细的方案，将 DMR 技术应用于检测 G 蛋白偶联受体的功能行为，该受体通常通过检测单个第二信使（如 cAMP、Ca(2+)或肌醇磷酸盐）的终点分析来进行检查。该方法可以轻松适应各种细胞背景（贴壁或悬浮），包括原代人细胞。实时记录可以在 384 孔微量滴定板中进行，2 小时内完成，也可以根据要解决的生物学问题延长至数小时。整个过程，包括细胞收获和 DMR 检测，需要 1-2 天。
