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Author

Soper, Steven A.
Soper Research Group
Research Interests
  • Micro- and nano-fabrication

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Polymer-based dense fluidic networks for high throughput screening with ultrasensitive fluorescence detection

Cited 4 times inthomson ciCited 4 times inthomson ci
Title
Polymer-based dense fluidic networks for high throughput screening with ultrasensitive fluorescence detection
Author
Okagbare, Paul I.Soper, Steven A.
Keywords
Enzyme activity; High throughput screening; Microfluidics; Ultrasensitive fluorescence
Issue Date
201009
Publisher
WILEY-V C H VERLAG GMBH
Citation
ELECTROPHORESIS, v.31, no.18, pp.3074 - 3082
Abstract
Microfluidics represents a viable platform for performing high throughput screening (HTS) because of its ability to automate fluid handling and generate fluidic networks with high number densities over small footprints appropriate for the simultaneous optical interrogation of many screening assays. While most HTS campaigns depend on fluorescence, readers typically use point detection and serially address the assay results significantly lowering throughput or detection sensitivity due to a low duty cycle. To address this challenge, we present here the fabrication of a high-density microfluidic network packed into the imaging area of a large field-of-view (FoV) ultrasensitive fluorescence detection system. The fluidic channels were 1, 5 or 10 mu m (width), 1 mu m (depth) with a pitch of 1-10 mu m and each fluidic processor was individually addressable. The fluidic chip was produced from a molding tool using hot embossing and thermal fusion bonding to enclose the fluidic channels. A 40 x microscope objective (numerical aperture = 0.75) created an FoV of 200 mu m, providing the ability to interrogate similar to 25 channels using the current fluidic configuration. An ultrasensitive fluorescence detection system with a large FoV was used to transduce fluorescence signals simultaneously from each fluidic processor onto the active area of an electron multiplying charge-coupled device. The utility of these multichannel networks for HTS was demonstrated by carrying out the high throughput monitoring of the activity of an enzyme, apurinic Endonuclease 1, used as a model-screening assay.
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DOI
http://dx.doi.org/10.1002/elps.201000209
ISSN
0173-0835
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