Single-Molecule Counting of Nucleotide by Electrophoresis with Nanochannel-Integrated Nano-Gap Devices
We utilized electrophoresis to control the fluidity of sample biomolecules in sample aqueous solutions inside the nanochannel for single-molecule detection by using a nanochannel-integrated nanogap electrode, which is composed of a nano-gap sensing electrode, nanochannel, and tapered focusing channe...
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doaj-a06ce55f0a01492597a769d280b8dcc02020-11-25T04:08:11ZengMDPI AGMicromachines2072-666X2020-10-011198298210.3390/mi11110982Single-Molecule Counting of Nucleotide by Electrophoresis with Nanochannel-Integrated Nano-Gap DevicesTakahito Ohshiro0Yuki Komoto1Masateru Taniguchi2Institute of Science and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka 567-0047, JapanInstitute of Science and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka 567-0047, JapanInstitute of Science and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka 567-0047, JapanWe utilized electrophoresis to control the fluidity of sample biomolecules in sample aqueous solutions inside the nanochannel for single-molecule detection by using a nanochannel-integrated nanogap electrode, which is composed of a nano-gap sensing electrode, nanochannel, and tapered focusing channel. In order to suppress electro-osmotic flow and thermal convection inside this nanochannel, we optimized the reduction ratios of the tapered focusing channel, and the ratio of inlet 10 μm to outlet 0.5 μm was found to be high performance of electrophoresis with lower concentration of 0.05 × TBE (Tris/Borate/EDTA) buffer containing a surfactant of 0.1 <i>w/v</i>% polyvinylpyrrolidone (PVP). Under the optimized conditions, single-molecule electrical measurement of deoxyguanosine monophosphate (dGMP) was performed and it was found that the throughput was significantly improved by nearly an order of magnitude compared to that without electrophoresis. In addition, it was also found that the long-duration signals that could interfere with discrimination were significantly reduced. This is because the strong electrophoresis flow inside the nanochannels prevents the molecules’ adsorption near the electrodes. This single-molecule electrical measurement with nanochannel-integrated nano-gap electrodes by electrophoresis significantly improved the throughput of signal detection and identification accuracy.https://www.mdpi.com/2072-666X/11/11/982nanochannelDNAsingle-molecule detection |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Takahito Ohshiro Yuki Komoto Masateru Taniguchi |
spellingShingle |
Takahito Ohshiro Yuki Komoto Masateru Taniguchi Single-Molecule Counting of Nucleotide by Electrophoresis with Nanochannel-Integrated Nano-Gap Devices Micromachines nanochannel DNA single-molecule detection |
author_facet |
Takahito Ohshiro Yuki Komoto Masateru Taniguchi |
author_sort |
Takahito Ohshiro |
title |
Single-Molecule Counting of Nucleotide by Electrophoresis with Nanochannel-Integrated Nano-Gap Devices |
title_short |
Single-Molecule Counting of Nucleotide by Electrophoresis with Nanochannel-Integrated Nano-Gap Devices |
title_full |
Single-Molecule Counting of Nucleotide by Electrophoresis with Nanochannel-Integrated Nano-Gap Devices |
title_fullStr |
Single-Molecule Counting of Nucleotide by Electrophoresis with Nanochannel-Integrated Nano-Gap Devices |
title_full_unstemmed |
Single-Molecule Counting of Nucleotide by Electrophoresis with Nanochannel-Integrated Nano-Gap Devices |
title_sort |
single-molecule counting of nucleotide by electrophoresis with nanochannel-integrated nano-gap devices |
publisher |
MDPI AG |
series |
Micromachines |
issn |
2072-666X |
publishDate |
2020-10-01 |
description |
We utilized electrophoresis to control the fluidity of sample biomolecules in sample aqueous solutions inside the nanochannel for single-molecule detection by using a nanochannel-integrated nanogap electrode, which is composed of a nano-gap sensing electrode, nanochannel, and tapered focusing channel. In order to suppress electro-osmotic flow and thermal convection inside this nanochannel, we optimized the reduction ratios of the tapered focusing channel, and the ratio of inlet 10 μm to outlet 0.5 μm was found to be high performance of electrophoresis with lower concentration of 0.05 × TBE (Tris/Borate/EDTA) buffer containing a surfactant of 0.1 <i>w/v</i>% polyvinylpyrrolidone (PVP). Under the optimized conditions, single-molecule electrical measurement of deoxyguanosine monophosphate (dGMP) was performed and it was found that the throughput was significantly improved by nearly an order of magnitude compared to that without electrophoresis. In addition, it was also found that the long-duration signals that could interfere with discrimination were significantly reduced. This is because the strong electrophoresis flow inside the nanochannels prevents the molecules’ adsorption near the electrodes. This single-molecule electrical measurement with nanochannel-integrated nano-gap electrodes by electrophoresis significantly improved the throughput of signal detection and identification accuracy. |
topic |
nanochannel DNA single-molecule detection |
url |
https://www.mdpi.com/2072-666X/11/11/982 |
work_keys_str_mv |
AT takahitoohshiro singlemoleculecountingofnucleotidebyelectrophoresiswithnanochannelintegratednanogapdevices AT yukikomoto singlemoleculecountingofnucleotidebyelectrophoresiswithnanochannelintegratednanogapdevices AT masaterutaniguchi singlemoleculecountingofnucleotidebyelectrophoresiswithnanochannelintegratednanogapdevices |
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