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Sensors 2014, 14(8), 15400-15414; doi:10.3390/s140815400

Rapid, Single-Molecule Assays in Nano/Micro-Fluidic Chips with Arrays of Closely Spaced Parallel Channels Fabricated by Femtosecond Laser Machining

Center for Laser Applications, University of Tennessee Space Institute, 411 B. H. Goethert Parkway, MS 35, Tullahoma, TN 37388, USA
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Received: 16 July 2014 / Revised: 8 August 2014 / Accepted: 18 August 2014 / Published: 20 August 2014
(This article belongs to the Special Issue Opto-Microfluidics for Bio Applications)
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Abstract

Cost-effective pharmaceutical drug discovery depends on increasing assay throughput while reducing reagent needs. To this end, we are developing an ultrasensitive, fluorescence-based platform that incorporates a nano/micro-fluidic chip with an array of closely spaced channels for parallelized optical readout of single-molecule assays. Here we describe the use of direct femtosecond laser machining to fabricate several hundred closely spaced channels on the surfaces of fused silica substrates. The channels are sealed by bonding to a microscope cover slip spin-coated with a thin film of poly(dimethylsiloxane). Single-molecule detection experiments are conducted using a custom-built, wide-field microscope. The array of channels is epi-illuminated by a line-generating red diode laser, resulting in a line focus just a few microns thick across a 500 micron field of view. A dilute aqueous solution of fluorescently labeled biomolecules is loaded into the device and fluorescence is detected with an electron-multiplying CCD camera, allowing acquisition rates up to 7 kHz for each microchannel. Matched digital filtering based on experimental parameters is used to perform an initial, rapid assessment of detected fluorescence. More detailed analysis is obtained through fluorescence correlation spectroscopy. Simulated fluorescence data is shown to agree well with experimental values. View Full-Text
Keywords: high-throughput; microfluidic; rapid readout; fluorescence correlation spectroscopy; femtosecond laser machining high-throughput; microfluidic; rapid readout; fluorescence correlation spectroscopy; femtosecond laser machining
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This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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MDPI and ACS Style

Canfield, B.K.; King, J.K.; Robinson, W.N.; Hofmeister, W.H.; Davis, L.M. Rapid, Single-Molecule Assays in Nano/Micro-Fluidic Chips with Arrays of Closely Spaced Parallel Channels Fabricated by Femtosecond Laser Machining. Sensors 2014, 14, 15400-15414.

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