Labelfree Technologiespast And Present

GPCR stimulation by agonists activates a variety of intracellular pathways via associated heterotrimeric G proteins and ß-arrestin signaling. These molecular events are mediated through specific interactions with downstream effector molecules, which ultimately lead to changes in cell physiology, such as cell adhesion, cell migration, cell proliferation, or even neurite outgrowth. A variety of technologies are used to measure either direct cell behavior or surrogate end points in response to GPCR stimulation in a quantitative and, occasionally, semiquantitative way. For example, cell migration can be measured in modified Boyden chambers in low-throughput to medium-throughput formats, or cell proliferation can be quantified using end point assays and surrogate markers such as DNA synthesis fH-thymidine, BrdU, DNA dye staining) or mitochondrial activity (MTT). Unfortunately, most of these traditional assay methods are laborious and frequently involve the use of radioactivity or dyes. It is therefore a challenge to develop label-free assay systems that allow precise monitoring of physiological changes of cells in response to ligand stimulation.

During the past years, several assay platforms and instruments were introduced to allow label-free, quantitative, and even real-time measurements of complex cell responses. The majority of these novel technologies find general and widespread applications in multiple areas of cell biology, but some of them, including microphysiometry, impedance, and resonant waveguide grating (RWG), are of special interest to the GPCR research field as they potentially offer novel information on GPCR pathway signatures and their consequences in cell physiology. For this particular reason, we elaborate here in particular on novel GPCR-relevant assay technologies.

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