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Electrokinetic manipulation of particles and cells in microfluidic devices.

机译:微流控设备中颗粒和细胞的电动操纵。

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摘要

With the recent advancement in micro-fabrication technology, lab-on-a-chip devices have been developed in order to perform biological analysis through cell manipulation. Microchannels used in these lab-on-a-chip devices have been demonstrated to accurately perform many different cell manipulation techniques such as focusing, separation, trapping, and lysis. Although there are many methods available for these techniques, electrokinetics has been rapidly gaining popularity due to the simplicity of application and removal of the need for in channel micro-structures. This thesis studies the use of electrokinetic flow and accompanying phenomena in various structured microchannels to perform focusing, separation, trapping, and lysis of cells. Three related projects were conducted in series.;First, a parametric study of the focusing of yeast cells using negative dielectrophoresis in a serpentine microchannel was studied. Focusing cells into a single stream is usually a necessary step prior to counting and separating them in microfluidic devices such as flow cytometers and cell sorters. This work demonstrated a sheathless electrokinetic focusing of yeast cells in a planar serpentine microchannel using DC-biased AC electric fields. The concurrent pumping and focusing of yeast cells arose from the DC electrokinetic transport and the turn-induced AC/DC dielectrophoretic motion, respectively. The effects of electric field (including AC to DC field ratio, and AC field frequency) and concentration (including buffer concentration and cell concentration) on the cell focusing performance were studied experimentally and numerically. A continuous electrokinetic filtration of E. coli cells from yeast cells was also demonstrated via their differential electrokinetic focusing in the serpentine microchannel.;Next, negative and positive dielectrophoretic focusing were also studied in their application to particle separation in a serpentine microchannel. This work first demonstrated negative and positive dielectrophoretic focusing of by changing only the electric conductivity of the suspending fluid. Due to the channel turn-induced dielectrophoretic force, particles were focused to either the centerline or the sidewalls of the channel when their electric conductivity was lower (i.e., negative DEP) or higher (i.e., positive DEP) than that of the fluid. These distinctive dielectrophoretic focusing phenomena in the serpentine microchannel were then combined to implement a continuous separation between particles of different sizes and electric conductivities. Such separation eliminates the fabrication of in-channel microelectrodes or micro-insulators that are typically required in DEP-based separation techniques.;Lastly, red blood cells were used to study cell lysis and trapping in a microchannel constriction. Cell Lysis is an important step in the analysis of intracellular contents. Electrical lysis of red blood cells was demonstrated in a hurdle microchannel using a low continuous DC-biased AC electric field amplified by channel geometry. Trapping of cells was also demonstrated using this DC-biased AC electric field, and the transition between trapping and lysis of red blood cells in this microchannel was demonstrated by simply adjusting the applied DC voltage. Further, these phenomena were used in conjunction to demonstrate the separation of Leukemia cells from red blood cells.
机译:随着微制造技术的最新发展,已经开发了芯片实验室设备,以便通过细胞操作进行生物学分析。这些芯片实验室设备中使用的微通道已被证明可以准确执行许多不同的细胞操作技术,例如聚焦,分离,捕获和裂解。尽管有许多方法可用于这些技术,但是由于应用的简便性和对通道微结构的需求的消除,电动动力学已迅速普及。本文研究了在各种结构化微通道中电动流及其伴随现象的使用,以进行细胞的聚焦,分离,捕获和裂解。连续进行了三个相关项目:首先,研究了在蛇形微通道中使用负介电电泳对酵母细胞聚焦的参数研究。在微流控设备(例如流式细胞仪和细胞分选仪)中对细胞进行计数和分离之前,通常需要将细胞集中到单个流中。这项工作证明了使用直流偏置交流电场在平面蛇形微通道中对酵母细胞进行无鞘电动聚焦。酵母细胞的同时泵送和聚焦分别来自直流电动运动和转向诱导的交流/直流介电泳运动。实验和数值研究了电场(包括交直流场比,交流场频率)和浓度(包括缓冲液浓度和细胞浓度)对细胞聚焦性能的影响。还通过在蛇形微通道中的差异电动聚焦,证明了从酵母细胞中对大肠杆菌细胞进行连续电动过滤。接着,还研究了负和正介电泳聚焦在蛇形微通道中分离颗粒中的应用。这项工作首先通过仅改变悬浮液的电导率证明了负电正电泳聚焦。由于通道转弯引起的介电泳力,当颗粒的电导率比流体的电导率低(即,负DEP)或更高(即,正DEP)时,颗粒会集中在通道的中心线或侧壁上。然后将蛇形微通道中的这些独特的介电泳聚焦现象组合起来,以实现不同大小和电导率的颗粒之间的连续分离。这种分离消除了在基于DEP的分离技术中通常需要的通道内微电极或微绝缘体的制造。最后,红细胞被用于研究细胞裂解和微通道缩窄中的捕获。细胞裂解是细胞内内容物分析中的重要步骤。在跨栏微通道中使用通过通道几何形状放大的低连续DC偏置AC电场证明了红细胞的电裂解。还使用该直流偏置的交流电场演示了细胞的捕获,并且通过简单调节施加的直流电压来演示了该微通道中红细胞的捕获和裂解之间的过渡。此外,将这些现象一起用于证明白血病细胞与红细胞的分离。

著录项

  • 作者

    Church, Christopher S.;

  • 作者单位

    Clemson University.;

  • 授予单位 Clemson University.;
  • 学科 Engineering Mechanical.
  • 学位 M.S.
  • 年度 2010
  • 页码 79 p.
  • 总页数 79
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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