Cell to aperture interaction in Patch-Clamp Chips visualized by Fluorescence microscopy and Focused-Ion Beam sections

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DOIResolve DOI: http://doi.org/10.1002/bit.23127
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TypeArticle
Journal titleBiotechnology and Bioengineering
Volume108
Issue8
Pages19361941; # of pages: 6
Subjection channels; planar patch-clamp chip; gigaseal; fluorescence microscopy; focused ion beam; scanning electron microscopy
AbstractPatch-clamp is an important method to monitor the electrophysiological activity of cells and the role of pharmacological compounds on specific ion channel roteins. In recent years, planar patch-clamp chips have been developed as a higher throughput approach to the established glass-pipette method. However, proper conditions to optimize the high resistance cell-to-probe seals required to measure the small currents resulting from ion channel activity are still the subject of conjecture. Here, we report on the design of multiple-aperture (sieve) chips to rapidly facilitate assessment of cell-to-aperture interactions in statistically significant numbers. We propose a method to prescreen the quality of seals based on a dye loading protocol through apertures in the chip and subsequent evaluation with fluorescence confocal microscopy. We also show the first scanning electron micrograph of a focused ion beam section of a cell in a patch-clamp chip aperture.
Publication date
LanguageEnglish
AffiliationNRC Institute for Microstructural Sciences; NRC Institute for Biological Sciences; National Research Council Canada
Peer reviewedYes
NPARC number18074298
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Record identifier460ff7b3-d06c-495d-9314-2f2adc0684b7
Record created2011-06-23
Record modified2017-03-23
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