On-site processing of single chromosomal DNA molecules using optically driven microtools on a microfluidic workbench

Abstract We developed optically driven microtools for processing single biomolecules using a microfluidic workbench composed of a microfluidic platform that functions under an optical microscope. The optically driven microtools have enzymes immobilized on their surfaces, which catalyze chemical reac...

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Autores principales:	Akihito Masuda, Hidekuni Takao, Fusao Shimokawa, Kyohei Terao
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e9beb0e14ae341d6972226ba4f4156da
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spelling	oai:doaj.org-article:e9beb0e14ae341d6972226ba4f4156da2021-12-02T18:03:21ZOn-site processing of single chromosomal DNA molecules using optically driven microtools on a microfluidic workbench10.1038/s41598-021-87238-32045-2322https://doaj.org/article/e9beb0e14ae341d6972226ba4f4156da2021-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-87238-3https://doaj.org/toc/2045-2322Abstract We developed optically driven microtools for processing single biomolecules using a microfluidic workbench composed of a microfluidic platform that functions under an optical microscope. The optically driven microtools have enzymes immobilized on their surfaces, which catalyze chemical reactions for molecular processing in a confined space. Optical manipulation of the microtools enables them to be integrated with a microfluidic device for controlling the position, orientation, shape of the target sample. Here, we describe the immobilization of enzymes on the surface of microtools, the microfluidics workbench, including its microtool storage and sample positioning functions, and the use of this system for on-site cutting of single chromosomal DNA molecules. We fabricated microtools by UV lithography with SU-8 and selected ozone treatments for immobilizing enzymes. The microfluidic workbench has tool-stock chambers for tool storage and micropillars to trap and extend single chromosomal DNA molecules. The DNA cutting enzymes DNaseI and DNaseII were immobilized on microtools that were manipulated using optical tweezers. The DNaseI tool shows reliable cutting for on-site processing. This pinpoint processing provides an approach for analyzing chromosomal DNA at the single-molecule level. The flexibility of the microtool design allows for processing of various samples, including biomolecules and single cells.Akihito MasudaHidekuni TakaoFusao ShimokawaKyohei TeraoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Akihito Masuda Hidekuni Takao Fusao Shimokawa Kyohei Terao On-site processing of single chromosomal DNA molecules using optically driven microtools on a microfluidic workbench
description	Abstract We developed optically driven microtools for processing single biomolecules using a microfluidic workbench composed of a microfluidic platform that functions under an optical microscope. The optically driven microtools have enzymes immobilized on their surfaces, which catalyze chemical reactions for molecular processing in a confined space. Optical manipulation of the microtools enables them to be integrated with a microfluidic device for controlling the position, orientation, shape of the target sample. Here, we describe the immobilization of enzymes on the surface of microtools, the microfluidics workbench, including its microtool storage and sample positioning functions, and the use of this system for on-site cutting of single chromosomal DNA molecules. We fabricated microtools by UV lithography with SU-8 and selected ozone treatments for immobilizing enzymes. The microfluidic workbench has tool-stock chambers for tool storage and micropillars to trap and extend single chromosomal DNA molecules. The DNA cutting enzymes DNaseI and DNaseII were immobilized on microtools that were manipulated using optical tweezers. The DNaseI tool shows reliable cutting for on-site processing. This pinpoint processing provides an approach for analyzing chromosomal DNA at the single-molecule level. The flexibility of the microtool design allows for processing of various samples, including biomolecules and single cells.
format	article
author	Akihito Masuda Hidekuni Takao Fusao Shimokawa Kyohei Terao
author_facet	Akihito Masuda Hidekuni Takao Fusao Shimokawa Kyohei Terao
author_sort	Akihito Masuda
title	On-site processing of single chromosomal DNA molecules using optically driven microtools on a microfluidic workbench
title_short	On-site processing of single chromosomal DNA molecules using optically driven microtools on a microfluidic workbench
title_full	On-site processing of single chromosomal DNA molecules using optically driven microtools on a microfluidic workbench
title_fullStr	On-site processing of single chromosomal DNA molecules using optically driven microtools on a microfluidic workbench
title_full_unstemmed	On-site processing of single chromosomal DNA molecules using optically driven microtools on a microfluidic workbench
title_sort	on-site processing of single chromosomal dna molecules using optically driven microtools on a microfluidic workbench
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/e9beb0e14ae341d6972226ba4f4156da
work_keys_str_mv	AT akihitomasuda onsiteprocessingofsinglechromosomaldnamoleculesusingopticallydrivenmicrotoolsonamicrofluidicworkbench AT hidekunitakao onsiteprocessingofsinglechromosomaldnamoleculesusingopticallydrivenmicrotoolsonamicrofluidicworkbench AT fusaoshimokawa onsiteprocessingofsinglechromosomaldnamoleculesusingopticallydrivenmicrotoolsonamicrofluidicworkbench AT kyoheiterao onsiteprocessingofsinglechromosomaldnamoleculesusingopticallydrivenmicrotoolsonamicrofluidicworkbench
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On-site processing of single chromosomal DNA molecules using optically driven microtools on a microfluidic workbench

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