Title: Microfabrication of PMMA sensor chips for an incubation-type planar patch clamp

Authors: Noriko Takada; Masaki Aoyama; Mitsukazu Suzui; Yosuke Hachisu; Hitoshi Ohmori; Zhi-Hong Wang; Hidetaka Uno; Obuliraj Senthil Kumar; Yasutaka Nagaoka; Miho Goto-Satoh; Tsuneo Urisu

Addresses: Institute for Molecular Science, Myodaiji, Okazaki, 444-8585, Japan ' Institute for Molecular Science, Myodaiji, Okazaki, 444-8585, Japan ' Institute for Molecular Science, Myodaiji, Okazaki, 444-8585, Japan ' RIKEN, 2-1, Hirosawa, Wako-shi, Saitama, 351-0198, Japan ' RIKEN, 2-1, Hirosawa, Wako-shi, Saitama, 351-0198, Japan ' FIRST Research Center for Innovative Nanobiodevices, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan ' FIRST Research Center for Innovative Nanobiodevices, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan ' FIRST Research Center for Innovative Nanobiodevices, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan ' FIRST Research Center for Innovative Nanobiodevices, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan ' FIRST Research Center for Innovative Nanobiodevices, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan ' FIRST Research Center for Innovative Nanobiodevices, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan

Abstract: A sensor chip for use in an ion channel biosensor based on an incubation-type planar patch clamp was formed using both side hot embossing and focused ion beam (FIB) processing. A biosensor was constructed using the PMMA chip, and the performance of this chip was investigated by measuring the laser-stimulated ion channel current of a channel rhodopsin wide receiver (ChRWR) expressed on HEK293 cells. The observed current profile and membrane potential dependences agreed well with the data measured using a pipette patch clamp. The noise level of the channel current recorded using a 1 kHz low pass filter was 7 pA (rms). The sensor chip fabrication processes are readily modified to permit multi-point measurements; therefore, the excellent performance of the PMMA chip indicates that the chip is suitable for use in high-throughput ion channel biosensor screening applications.

Keywords: microfabrication; hot embossing; focused ion beam; FIB; ultra-precision machining; planar patch clamp; ion channel; biosensors; HEK293; polymethyl methacrylate; PMMA; nanomanufacturing; nanotechnology.

DOI: 10.1504/IJNM.2014.060798

International Journal of Nanomanufacturing, 2014 Vol.10 No.3, pp.281 - 294

Received: 31 Oct 2012
Accepted: 19 Jul 2013
Published online: 17 May 2014 *

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