Electrokinetic Delivery of Single Fluorescent Biomolecules in Fluidic Nanochannels
Abstract
We describe the fabrication of sub-100-nanometer-sized channels in a fused silica lab-on-a-chip device and experiments that demonstrate detection of single fluorescently labeled proteins in buffer solution within the device with high signal and low background. The fluorescent biomolecules are transported along the length of the nanochannels by electrophoresis and/or electro-osmosis until they pass into a two-focus laser irradiation zone. Pulse-interleaved excitation and time-resolved single-photon detection with maximum-likelihood analysis enables the location of the biomolecule to be determined. Diffusional transport of the molecules is found to be slowed within the nanochannel, and this facilitates fluidic trapping and/or prolonged measurements on individual biomolecules. Our goal is to actively control the fluidic transport to achieve rapid delivery of each new biomolecule to the sensing zone, following the completion of measurements, or the photobleaching of the prior molecule. We have used computer simulations that include photophysical effects such as triplet crossing and photobleaching of the labels to design control algorithms, which are being implemented in a custom field-programmable-gate-array circuit for the active fluidic control.
- Authors:
-
- University of Tennessee Space Institute
- Vanderbilt University
- ORNL
- Publication Date:
- Research Org.:
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Center for Nanophase Materials Sciences (CNMS); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Shared Research Equipment Collaborative Research Center
- Sponsoring Org.:
- Work for Others (WFO)
- OSTI Identifier:
- 965844
- DOE Contract Number:
- DE-AC05-00OR22725
- Resource Type:
- Conference
- Resource Relation:
- Conference: SPIE NanoScience + Engineering 2008, San Diego, CA, USA, 20080810, 20080814
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; ALGORITHMS; BUFFERS; COMPUTERIZED SIMULATION; DESIGN; DETECTION; ELECTRODYNAMICS; ELECTROPHORESIS; EXCITATION; FABRICATION; IRRADIATION; LASERS; PROTEINS; SILICA; TRANSPORT; TRAPPING; TRIPLETS
Citation Formats
Davis, Lloyd M, Canfield, Brian K, Li, Xiaoxuan, Hofmeister, William, Shen, Guoqing, Lescano, Isaac, Bomar, Bruce W, Wikswo, John P, Markov, Dmitry P, Samson, Philip C, Daniel, Claus, Sikorski, Zbigniew, and Robinson, William N. Electrokinetic Delivery of Single Fluorescent Biomolecules in Fluidic Nanochannels. United States: N. p., 2008.
Web.
Davis, Lloyd M, Canfield, Brian K, Li, Xiaoxuan, Hofmeister, William, Shen, Guoqing, Lescano, Isaac, Bomar, Bruce W, Wikswo, John P, Markov, Dmitry P, Samson, Philip C, Daniel, Claus, Sikorski, Zbigniew, & Robinson, William N. Electrokinetic Delivery of Single Fluorescent Biomolecules in Fluidic Nanochannels. United States.
Davis, Lloyd M, Canfield, Brian K, Li, Xiaoxuan, Hofmeister, William, Shen, Guoqing, Lescano, Isaac, Bomar, Bruce W, Wikswo, John P, Markov, Dmitry P, Samson, Philip C, Daniel, Claus, Sikorski, Zbigniew, and Robinson, William N. 2008.
"Electrokinetic Delivery of Single Fluorescent Biomolecules in Fluidic Nanochannels". United States.
@article{osti_965844,
title = {Electrokinetic Delivery of Single Fluorescent Biomolecules in Fluidic Nanochannels},
author = {Davis, Lloyd M and Canfield, Brian K and Li, Xiaoxuan and Hofmeister, William and Shen, Guoqing and Lescano, Isaac and Bomar, Bruce W and Wikswo, John P and Markov, Dmitry P and Samson, Philip C and Daniel, Claus and Sikorski, Zbigniew and Robinson, William N},
abstractNote = {We describe the fabrication of sub-100-nanometer-sized channels in a fused silica lab-on-a-chip device and experiments that demonstrate detection of single fluorescently labeled proteins in buffer solution within the device with high signal and low background. The fluorescent biomolecules are transported along the length of the nanochannels by electrophoresis and/or electro-osmosis until they pass into a two-focus laser irradiation zone. Pulse-interleaved excitation and time-resolved single-photon detection with maximum-likelihood analysis enables the location of the biomolecule to be determined. Diffusional transport of the molecules is found to be slowed within the nanochannel, and this facilitates fluidic trapping and/or prolonged measurements on individual biomolecules. Our goal is to actively control the fluidic transport to achieve rapid delivery of each new biomolecule to the sensing zone, following the completion of measurements, or the photobleaching of the prior molecule. We have used computer simulations that include photophysical effects such as triplet crossing and photobleaching of the labels to design control algorithms, which are being implemented in a custom field-programmable-gate-array circuit for the active fluidic control.},
doi = {},
url = {https://www.osti.gov/biblio/965844},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Tue Jan 01 00:00:00 EST 2008},
month = {Tue Jan 01 00:00:00 EST 2008}
}