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Authored by: Paul Taylor, Boehringer Ingleheim Inc.
This technique is an alternative to patch clamp electrophysiology for measuring changes in membrane potential upon activation of ion channels. Rubidium is similar to K+ in its capability of penetrating ion channels and, because of its absence in eukaryotic cells, avoids the possibility of endogenously produced false signals. Cells under study have their culturing buffer containing K+ exchanged out with a buffer containing Rb+ which initiates a cell-loading phase. Following the loading stage, excess Rb+ is washed away in preparation for ion channel activation. This is accomplished either by adding a depolarizing concentration of KCl (voltage-gated) or an appropriate ligand (ligand-gated). Cell supernatants containing effluxed rubidium are then analyzed using atomic absorption spectrometry (AAS) and instrumentation has been developed that allows throughputs of up to 60,000 samples per day using 384-well plates.
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