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Ca2+ ion transport through channels formed by α-hemolysin analyzed using a microwell array on a Si substrate
- Source :
-
Biosensors & Bioelectronics . Jan2012, Vol. 31 Issue 1, p445-450. 6p. - Publication Year :
- 2012
-
Abstract
- Abstract: For the functional analysis of ion channel activity, an artificial lipid bilayer suspended over microwells was formed that ruptured giant unilamellar vesicles on a Si substrate. Ca2+ ion indicators (fluo-4) were confined in the microwells by sealing the microwells with a lipid bilayer. An overhang formed at the microwells prevented the lipid membrane from falling into them and allowed the stable confinement of the fluorescent probes. The transport of Ca2+ ions through the channels formed by α-hemolysin inserted in a lipid membrane was analyzed by employing the fluorescence intensity change of fluo-4 in the microwells. The microwell volume was very small (1–100fl), so a highly sensitive monitor could be realized. The detection limit is several tens of ions/s/μm2, and this is much smaller than the ion current in a standard electrophysiological measurement. Smaller microwells will make it possible to mimic a local ion concentration change in the cells, although the signal to noise ratio must be further improved for the functional analysis of a single channel. We demonstrated that a microwell array with confined fluorescent probes sealed by a lipid bilayer could constitute a basic component of a highly sensitive biosensor array that works with functional membrane proteins. This array will allow us to realize high throughput and parallel testing devices. [Copyright &y& Elsevier]
Details
- Language :
- English
- ISSN :
- 09565663
- Volume :
- 31
- Issue :
- 1
- Database :
- Academic Search Index
- Journal :
- Biosensors & Bioelectronics
- Publication Type :
- Academic Journal
- Accession number :
- 70157161
- Full Text :
- https://doi.org/10.1016/j.bios.2011.11.010