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

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 optica...

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Autores principales: Canfield, Brian K., King, Jason K., Robinson, William N., Hofmeister, William H., Davis, Lloyd M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4178987/
https://www.ncbi.nlm.nih.gov/pubmed/25140634
http://dx.doi.org/10.3390/s140815400
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author Canfield, Brian K.
King, Jason K.
Robinson, William N.
Hofmeister, William H.
Davis, Lloyd M.
author_facet Canfield, Brian K.
King, Jason K.
Robinson, William N.
Hofmeister, William H.
Davis, Lloyd M.
author_sort Canfield, Brian K.
collection PubMed
description 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.
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spelling pubmed-41789872014-10-02 Rapid, Single-Molecule Assays in Nano/Micro-Fluidic Chips with Arrays of Closely Spaced Parallel Channels Fabricated by Femtosecond Laser Machining Canfield, Brian K. King, Jason K. Robinson, William N. Hofmeister, William H. Davis, Lloyd M. Sensors (Basel) Article 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. MDPI 2014-08-20 /pmc/articles/PMC4178987/ /pubmed/25140634 http://dx.doi.org/10.3390/s140815400 Text en © 2014 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Canfield, Brian K.
King, Jason K.
Robinson, William N.
Hofmeister, William H.
Davis, Lloyd M.
Rapid, Single-Molecule Assays in Nano/Micro-Fluidic Chips with Arrays of Closely Spaced Parallel Channels Fabricated by Femtosecond Laser Machining
title Rapid, Single-Molecule Assays in Nano/Micro-Fluidic Chips with Arrays of Closely Spaced Parallel Channels Fabricated by Femtosecond Laser Machining
title_full Rapid, Single-Molecule Assays in Nano/Micro-Fluidic Chips with Arrays of Closely Spaced Parallel Channels Fabricated by Femtosecond Laser Machining
title_fullStr Rapid, Single-Molecule Assays in Nano/Micro-Fluidic Chips with Arrays of Closely Spaced Parallel Channels Fabricated by Femtosecond Laser Machining
title_full_unstemmed Rapid, Single-Molecule Assays in Nano/Micro-Fluidic Chips with Arrays of Closely Spaced Parallel Channels Fabricated by Femtosecond Laser Machining
title_short Rapid, Single-Molecule Assays in Nano/Micro-Fluidic Chips with Arrays of Closely Spaced Parallel Channels Fabricated by Femtosecond Laser Machining
title_sort rapid, single-molecule assays in nano/micro-fluidic chips with arrays of closely spaced parallel channels fabricated by femtosecond laser machining
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4178987/
https://www.ncbi.nlm.nih.gov/pubmed/25140634
http://dx.doi.org/10.3390/s140815400
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