Grad Michael, Bigelow Alan W, Garty Guy, Attinger Daniel, Brenner David J
Department of Mechanical Engineering, Columbia University, New York, New York 10027, USA.
Rev Sci Instrum. 2013 Jan;84(1):014301. doi: 10.1063/1.4774043.
This paper describes the fabrication and integration of light-induced dielectrophoresis for cellular manipulation in biological microbeams. An optoelectronic tweezers (OET) cellular manipulation platform was designed, fabricated, and tested at Columbia University's Radiological Research Accelerator Facility (RARAF). The platform involves a light induced dielectrophoretic surface and a microfluidic chamber with channels for easy input and output of cells. The electrical conductivity of the particle-laden medium was optimized to maximize the dielectrophoretic force. To experimentally validate the operation of the OET device, we demonstrate UV-microspot irradiation of cells containing green fluorescent protein (GFP) tagged DNA single-strand break repair protein, targeted in suspension. We demonstrate the optofluidic control of single cells and groups of cells before, during, and after irradiation. The integration of optofluidic cellular manipulation into a biological microbeam enhances the facility's ability to handle non-adherent cells such as lymphocytes. To the best of our knowledge, this is the first time that OET cell handling is successfully implemented in a biological microbeam.
本文描述了用于生物微束中细胞操控的光诱导介电泳的制造与集成。在哥伦比亚大学放射研究加速器设施(RARAF)设计、制造并测试了一个光电镊子(OET)细胞操控平台。该平台包括一个光诱导介电泳表面和一个带有通道的微流体腔室,便于细胞的输入和输出。对含有颗粒的介质的电导率进行了优化,以最大化介电泳力。为了通过实验验证OET装置的运行,我们展示了对悬浮状态下含有绿色荧光蛋白(GFP)标记的DNA单链断裂修复蛋白的细胞进行紫外微斑照射。我们展示了在照射前、照射期间和照射后对单细胞和细胞群的光流体控制。将光流体细胞操控集成到生物微束中增强了该设施处理诸如淋巴细胞等非贴壁细胞的能力。据我们所知,这是首次在生物微束中成功实现OET细胞操控。