Silicon nanopore patch clamp device for measuring single ion channel activity

Plucinski, Lisa

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https://hdl.handle.net/2142/73092 Copy

Description

Title

Silicon nanopore patch clamp device for measuring single ion channel activity

Author(s)

Plucinski, Lisa

Issue Date

2015-01-21

Director of Research (if dissertation) or Advisor (if thesis)

Liu, Gang Logan

Department of Study

Bioengineering

Discipline

Bioengineering

Degree Granting Institution

University of Illinois at Urbana-Champaign

Degree Name

M.S.

Degree Level

Thesis

Keyword(s)

• Nanopore
• Nanoporous
• Patch clamp
• SH-SY5Y

Abstract

The development of new methods for measuring and manipulating neural activity is of high interest with applications in neuron and neural circuit functioning as well as high-throughput ion channel screening for early drug development research. In 1991 Erwin Neher and Bert Sakmann received the Nobel Prize for their invention of the patch clamp, which is based on the formation of a high resistance seal between a glass micropipette and a cell membrane. This transformative method allows low noise recording of current arising from single ion channels or from an entire cell and has no equivalent in experimental neuroscience. However, the consistent formation and maintenance of a high resistance seal is nontrivial and unpredictable, requiring a large amount of time and labor, which is inconvenient and limits applications. In order to automate the patch clamp process, researchers have developed patch clamp on chip devices that are optimized for whole cell recordings. Here we have developed a novel silicon nanopore planar patch clamp chip that is ideal for single ion channel measurements because the nanopore apertures minimize cell membrane capacitance, reduce chip capacitance, reduce cell membrane damage, and allow for work with smaller cells. Here we give an overview of nanopore and nanoporous biodevices, the traditional patch clamp technqiue, requirements for noise reduction in patch clamp experiments, and recent advances in automated patch clamp. We then discuss our silicon nanopore fabrication method, which is based on wafer-scale anisotropic wet etching methods, and describe how this is incorporated into our patch clamp on chip device. We carry out electrical characterization of our devices before use to ensure that the impedance fulfills the requirements currently in place for single channel patch clamp recordings. In addition to device fabrication and characterization, we also present the single channel recordings we have obtained from the human neuron-like SH-SY5Y cell line cultured and differentiated on-chip.

Graduation Semester

2014-12

Permalink

http://hdl.handle.net/2142/73092

Copyright and License Information

Copyright 2014 Lisa Anne Plucinski

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