Simonnet Claire, Groisman Alex
Department of Physics, University of California, San Diego, 9500 Gilman Drive, La Jolla, California 92093, USA.
Anal Chem. 2006 Aug 15;78(16):5653-63. doi: 10.1021/ac060340o.
We describe the design, fabrication, and operation of two types of flow cytometers based on microfluidic devices made of a single cast of poly(dimethylsiloxane). The stream of particles or cells injected into the devices is hydrodynamically focused in both transverse and lateral directions, has a uniform velocity, and has adjustable diameter and shape. The cytometry system built around the first microfluidic device has fluorescence detection accuracy comparable with that of a commercial flow cytometer and can analyze as many as 17 000 particles/s. This high-throughput microfluidic device could be used in inexpensive stand-alone cytometers or as a part of integrated microanalysis systems. In the second device, a stream of particles is focused to a flow layer of a submicrometer thickness that allows imaging the particles with a high numerical aperture microscope objective. To take long-exposure, low-light fluorescence images of live cells, the device is placed on a moving stage, which accurately balances the translational motion of particles in the flow. The achieved resolution is comparable to that of still micrographs. This high-resolution device could be used for analysis of morphology and fluorescence distribution in cells in continuous flow.
我们描述了基于由单铸聚二甲基硅氧烷制成的微流控装置的两种流式细胞仪的设计、制造和操作。注入装置的颗粒或细胞流在横向和侧向都受到流体动力学聚焦，具有均匀的速度，并且直径和形状可调节。围绕第一个微流控装置构建的细胞计数系统具有与商业流式细胞仪相当的荧光检测精度，并且每秒可分析多达17000个颗粒。这种高通量微流控装置可用于廉价的独立细胞仪或作为集成微分析系统的一部分。在第二个装置中，颗粒流被聚焦到亚微米厚度的流动层，这允许使用高数值孔径显微镜物镜对颗粒进行成像。为了获取活细胞的长时间曝光、低光荧光图像，该装置放置在移动平台上，该平台精确平衡了流动中颗粒的平移运动。所实现的分辨率与静态显微照片相当。这种高分辨率装置可用于分析连续流动细胞中的形态和荧光分布。
