Masuda Akihito, Takao Hidekuni, Shimokawa Fusao, Terao Kyohei
Department of Intelligent Mechanical Systems Engineering, Kagawa University, Takamatsu, 761-0396, Japan.
Nano-Micro Structure Device Integrated Research Center, Kagawa University, Takamatsu, 761-0396, Japan.
Sci Rep. 2021 Apr 12;11(1):7961. doi: 10.1038/s41598-021-87238-3.
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.
我们开发了用于处理单个生物分子的光驱动微工具，该工具使用由在光学显微镜下运行的微流控平台组成的微流控工作台。光驱动微工具的表面固定有酶，这些酶在受限空间内催化分子加工的化学反应。对微工具的光学操纵使其能够与微流控装置集成，以控制目标样品的位置、方向和形状。在此，我们描述了酶在微工具表面的固定、微流控工作台，包括其微工具存储和样品定位功能，以及该系统用于现场切割单个染色体DNA分子的应用。我们通过使用SU-8的紫外光刻和选定的臭氧处理来制造微工具，以固定酶。微流控工作台具有用于工具存储的工具库室和用于捕获和拉伸单个染色体DNA分子的微柱。DNA切割酶DNaseI和DNaseII被固定在使用光镊操纵的微工具上。DNaseI工具显示出用于现场加工的可靠切割效果。这种精确加工为在单分子水平分析染色体DNA提供了一种方法。微工具设计的灵活性允许处理各种样品，包括生物分子和单细胞。
