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A SIMPLE AND AFFORDABLE METHOD FOR IMMUNOPHENOTYPING USING A MICROFLUIDIC CHIP SAMPLE PREPARATION WITH IMAGE CYTOMETRY
A SIMPLE AND AFFORDABLE METHOD FOR IMMUNOPHENOTYPING USING A MICROFLUIDIC CHIP SAMPLE PREPARATION WITH IMAGE CYTOMETRY
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机译：图像细胞微流片芯片制备免疫印迹法的简便易行方法
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摘要
The enumeration of cells in fluids by flow cytometry is widely used across many disciplines such as assessment of leukocyte subsets in different bodily fluids or of bacterial contamination in environmental samples, food products and bodily fluids. For many applications the cost, size and complexity of the instruments prevents wider use, for example, CD4 analysis in HIV monitoring in resource-poor countries. The novel device, methods and system disclosed herein largely overcome these limitations. The system includes a simple system for CD4 and CD8 counting in point-of-care HIV staging within resource poor countries. Unlike previous approaches, no sample preparation is required with the sample added directly to a chip containing dried reagents by capillary flow. A large area image cytometer consisting of an LED module is used to excite the fluorochromes PerCP and APC labeled targets and a monochrome CCD camera with a combination of two macro lenses captures images of 40 mm2 of blood (approximately 1 microliter). CD4 and CD8-T-lymphocyte counts correlate well with those obtained by flow cytometry. The cytometer system described in the present invention provides an affordable and easy-to-use technique for use in remote locations.
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机译：通过流式细胞术对液体中的细胞进行计数已广泛用于许多学科，例如评估不同体液中的白细胞亚群或环境样品，食品和体液中的细菌污染。在许多应用中，这些工具的成本，规模和复杂性妨碍了其更广泛的使用，例如，资源贫乏国家的艾滋病毒监测中的CD4分析。本文公开的新颖的装置，方法和系统在很大程度上克服了这些限制。该系统包括一个简单的CD4和CD8系统，用于在资源匮乏的国家/地区对即时医疗艾滋病分期进行计数。与先前的方法不同，不需要通过直接将样品通过毛细管流添加到包含干燥试剂的芯片中的样品制备方法。由LED模块组成的大面积图像流式细胞仪用于激发荧光染料PerCP和APC标记的靶标，带有两个微距镜头的单色CCD摄像头可捕获40 mm 2 的血液图像（大约1微升）。 CD4和CD8-T淋巴细胞计数与通过流式细胞仪获得的计数非常相关。本发明中描述的细胞仪系统提供了一种在偏远地区使用的负担得起且易于使用的技术。
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