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一种用于细胞裂解和样品匀浆的新型光盘状离心微流控系统。

A novel, compact disk-like centrifugal microfluidics system for cell lysis and sample homogenization.

作者信息

Kido Horacio, Micic Miodrag, Smith David, Zoval Jim, Norton Jim, Madou Marc

机构信息

Department of Mechanical and Aerospace Engineering, University of California, Irvine, Irvine, CA 92697-3975, USA.

出版信息

Colloids Surf B Biointerfaces. 2007 Jul 1;58(1):44-51. doi: 10.1016/j.colsurfb.2007.03.015. Epub 2007 Mar 27.

Abstract

In this paper, we present the design and characterization of a novel platform for mechanical cell lysis of even the most difficult to lyse cell types on a micro or nanoscale (maximum 70 microL total volume). The system incorporates a machined plastic circular disk assembly, magnetic field actuated microfluidics, centrifugal cells and tissue homogenizer and centrifugation system. The mechanism of tissue disruption of this novel cell homogenization apparatus derives from the relative motion of ferromagnetic metal disks and grinding matrices in a liquid medium within individual chambers of the disk in the presence of an oscillating magnetic field. The oscillation of the ferromagnetic disks or blades produces mechanical impaction and shear forces capable of disrupting cells within the chamber both by direct action of the blade and by the motion of the surrounding lysis matrix, and by motion induced vortexing of buffer fluid. Glass beads or other grinding media are integrated into each lysis chamber within the disk to enhance the transfer of energy from the oscillating metal blade to the cells. The system also achieves the centrifugal elimination of solids from each liquid sample and allows the elution of clarified supernatants via siphoning into a collection chamber fabricated into the plastic disk assembly. This article describes system design, implementation and validation of proof of concept on two samples--Escherichia coli and Saccharomyces cerevisiae representing model systems for cells that are easy and difficult to lyse, respectively.

摘要

在本文中,我们展示了一种新型平台的设计与特性,该平台可在微纳尺度(总体积最大70微升)对即使是最难裂解的细胞类型进行机械细胞裂解。该系统包括一个加工成型的塑料圆盘组件、磁场驱动的微流体装置、离心式细胞和组织匀浆器以及离心系统。这种新型细胞匀浆设备的组织破坏机制源于在振荡磁场存在的情况下,铁磁金属圆盘与圆盘各个腔室内液体介质中的研磨基质之间的相对运动。铁磁圆盘或叶片的振荡会产生机械冲击和剪切力,这些力能够通过叶片的直接作用、周围裂解基质的运动以及缓冲液运动引起的涡旋,来破坏腔室内的细胞。玻璃珠或其他研磨介质被整合到圆盘内的每个裂解腔室中,以增强从振荡金属叶片到细胞的能量传递。该系统还能通过离心从每个液体样品中去除固体,并允许通过虹吸将澄清的上清液洗脱到塑料圆盘组件中制成的收集腔室。本文描述了该系统的设计、实现以及在两个样品(分别代表易于裂解和难以裂解的细胞模型系统的大肠杆菌和酿酒酵母)上的概念验证验证。

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