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基于手机的宽视野荧光显微镜和荧光成像流式细胞术。

Wide-field fluorescent microscopy and fluorescent imaging flow cytometry on a cell-phone.

作者信息

Zhu Hongying, Ozcan Aydogan

机构信息

Electrical Engineering Department, University of California, Los Angeles, USA.

出版信息

J Vis Exp. 2013 Apr 11(74):50451. doi: 10.3791/50451.

Abstract

Fluorescent microscopy and flow cytometry are widely used tools in biomedical research and clinical diagnosis. However these devices are in general relatively bulky and costly, making them less effective in the resource limited settings. To potentially address these limitations, we have recently demonstrated the integration of wide-field fluorescent microscopy and imaging flow cytometry tools on cell-phones using compact, light-weight, and cost-effective opto-fluidic attachments. In our flow cytometry design, fluorescently labeled cells are flushed through a microfluidic channel that is positioned above the existing cell-phone camera unit. Battery powered light-emitting diodes (LEDs) are butt-coupled to the side of this microfluidic chip, which effectively acts as a multi-mode slab waveguide, where the excitation light is guided to uniformly excite the fluorescent targets. The cell-phone camera records a time lapse movie of the fluorescent cells flowing through the microfluidic channel, where the digital frames of this movie are processed to count the number of the labeled cells within the target solution of interest. Using a similar opto-fluidic design, we can also image these fluorescently labeled cells in static mode by e.g. sandwiching the fluorescent particles between two glass slides and capturing their fluorescent images using the cell-phone camera, which can achieve a spatial resolution of e.g. - 10 μm over a very large field-of-view of - 81 mm(2). This cell-phone based fluorescent imaging flow cytometry and microscopy platform might be useful especially in resource limited settings, for e.g. counting of CD4+ T cells toward monitoring of HIV+ patients or for detection of water-borne parasites in drinking water.

摘要

荧光显微镜和流式细胞仪是生物医学研究和临床诊断中广泛使用的工具。然而,这些设备总体上相对笨重且昂贵,在资源有限的环境中效果较差。为了潜在地解决这些限制,我们最近展示了使用紧凑、轻便且经济高效的光流体附件,将宽场荧光显微镜和成像流式细胞仪工具集成到手机上。在我们的流式细胞仪设计中,荧光标记的细胞被冲洗通过位于现有手机摄像头单元上方的微流体通道。由电池供电的发光二极管(LED)与该微流体芯片的侧面对接耦合,该芯片有效地充当多模平板波导,激发光在其中被引导以均匀激发荧光靶标。手机摄像头记录流经微流体通道的荧光细胞的延时电影,对这部电影的数字帧进行处理以计算目标感兴趣溶液中标记细胞的数量。使用类似的光流体设计,我们还可以通过例如将荧光颗粒夹在两个载玻片之间并使用手机摄像头捕获其荧光图像,以静态模式对这些荧光标记的细胞进行成像,在 - 81 mm(2) 的非常大的视野上可以实现例如 - 10 μm 的空间分辨率。这种基于手机的荧光成像流式细胞术和显微镜平台可能特别适用于资源有限的环境,例如用于对艾滋病毒阳性患者进行监测时计数 CD4 + T 细胞,或用于检测饮用水中的水传播寄生虫。

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