کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
869453 909830 2007 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Integrated microsystem for non-invasive electrophysiological measurements on Xenopus oocytes
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Integrated microsystem for non-invasive electrophysiological measurements on Xenopus oocytes
چکیده انگلیسی

We propose a new non-invasive integrated microsystem for electrophysiological measurements on Xenopus laevis oocytes. Xenopus oocyte is a well-known expression system for various kinds of ion channels, that are potential tools in drug screening. In the traditional “Two Electrode Voltage Clamp” (TEVC) method, delicate micromanipulation is required to impale an oocyte with two microelectrodes. In our system, a non-invasive electrical access to the cytoplasm is provided by permeabilizing the cell membrane with an ionophore (e.g. nystatin). Unlike the classical patch-clamp or “macropatch” techniques, this method does not require removal of the vitelline membrane. Cell handling is significantly simplified, resulting in more robust recordings with increased throughput. Moreover, because only part of the oocyte surface is exposed to reagents, the required volume of reagent solutions could be reduced by an order of magnitude compared to the TEVC method. The fabrication process for this disposable microchip, based on poly-dimethylsiloxane (PDMS) molding and glass/PDMS bonding, is cost-efficient and simple. We tested this new microdevice by recording currents in oocytes expressing the human Epithelial Sodium Channel (hENaC) for membrane potentials between −100 and +50 mV. We recorded benzamil-sensitive currents with a large signal-to-noise ratio and we also obtained a benzamil concentration–inhibition curve displaying an inhibition constant IC50 of about 50 nM, comparable to previously published values obtained with the TEVC technique.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biosensors and Bioelectronics - Volume 22, Issue 12, 15 June 2007, Pages 3196–3202
نویسندگان
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