DNA Manipulation and Separation in Sublithographic-Scale Nanowire Array

Abstract

Electrokinetic manipulations of biomolecules using artificial nanostructures within microchannels have proven capability for controlling the dynamics of biomolecules. Because there is an inherent spatial size limitation to lithographic technology, especially for nanostructures with a small diameter and high aspect ratio, manipulating a single small biomolecule such as in DNA elongation before nanopore sequencing is still troublesome. Here we show the feasibility for self-assembly of a nanowire array embedded in a microchannel on a fused silica substrate as a means to manipulate the dynamics of a single long T4-DNA molecule and also separate DNA molecules. High-resolution optical microscopy measurements are used to clarify the presence of fully elongated T4-DNA molecules in the nanowire array. The spatial controllability of sublithographic-scale nanowires within microchannels offers a flexible platform not only for manipulating and separating long DNA molecules but also for integrating with other nanostructures to detect biomolecules in methods such as nanopore sequencing.

Exploiting biased reptation for continuous flow preparative DNA fractionation in a versatile microfluidic platform

“…The reservoir volume is 2000 times larger than the inner volume of the microchip and the buffer solution was frequently refreshed. The μGEL device has a calculated throughput of 0.18 ng of molecules per hour at the DNA input concentration of 12.5 ng μL − 1 and is comparable to previously reported microfabricated devices 20,31,32 .…”

Nano- and Microbiodevices for High-Performance Separation of Biomolecules

“…4, but it is hard to fabricate nanopillar less than 100-nm diameter, which is required to separate smaller molecules, including protein, sugar chain, lipid, and microRNA. Vapor-liquid-solid methods were utilized to fabricate nanowire structures, which are less than 100-nm diameter, inside microfabricated channels [28,29,33]. Even three-dimensional nanowire structures are possible to fabricate inside a microchannel as shown in Fig.…”

Nanobiodevice-based Single Biomolecule Analysis, Single-Cell Analysis, and in vivo Imaging for Cancer Diagnosis, Cancer Theranostics, and iPS Cell-based Regenerative Medicine

“…Two different types of nanofabrication technologies were established: 1) the so-called top-down nanotechnology, which is combined electron-beam lithography and plasma dry etching and 2) so-called bottom-up nanotechnology, which involves vapor-liquid-solid nanowire growth technique, to make nanopillar 2,5,6,41,46 and nanowire 56 structures with a high aspect ratio on quartz glass chips as shown in Fig. 2.…”

“…[16][17][18] The research efforts in my laboratory have been focused on the development of novel nanobiodevices for biomedical applications. [1][2][3][4][5][6][7] In this review article, we mainly overview our research activities, including nanobiodevices for single-cell analysis, [20][21][22][23][24][25] single biomolecule analysis, 19,[26][27][28][29]41,46,50,[54][55][56][57][58] biomarker detections, 23,[30][31][32][33]39,[47][48][49] cancer theranostics, and iPS cell in vivo imaging. [20][21][22][23][24][25][34][35][36][37][38]…”