Filter-based microfluidic device as a platform for immunofluorescent assay of microbial cells
Filter-based microfluidic device as a platform for immunofluorescent assay of microbial cells
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DOI：
10.1039/b401834f
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发表时间：
2004-01-01
期刊：
影响因子：
6.1
通讯作者：
Liu, WT
中科院分区：
文献类型：
--
作者：
Zhu, L;Zhang, Q;Liu, WT
基于过滤器的微流控装置与免疫荧光标记相结合，作为快速检测微生物细胞的平台。硬币大小的装置由微室、微通道和过滤堰（间隙= 1-2 μ m）组成，并且被证明能有效地捕获和浓缩微生物细胞（即，隐孢子虫和蓝氏贾第鞭毛虫），其尺寸大于堰隙。在样品注入后，将含有荧光标记抗体的染色溶液连续提供到装置中（流速= 20穆尔/分钟（-1）），以将微生物细胞冲向堰并加速荧光标记反应。使用比常规玻璃方法中使用的推荐浓度稀释10至100倍的染色溶液，可以在二次洗涤前2至5分钟内以单细胞水平显微镜观察荧光信噪比为12的那些靶细胞。
A filter-based microfluidic device was combined with immunofluorescent labeling as a platform to rapidly detect microbial cells. The coin-sized device consisted of micro-chambers, micro-channels and filter weirs (gap = 1-2 mum), and was demonstrated to effectively trap and concentrate microbial cells (i.e., Cryptosporidium parvum and Giardia lamblia), which were larger in size than the weir gap. After sample injection, a staining solution containing fluorescently-labeled antibodies was continuously provided into the device (flow rate = 20 mul min(-1)) to flush the microbial cells toward the weirs and to accelerate the fluorescent labeling reaction. Using a staining solution that was 10 to 100 times more dilute than the recommended concentration used in a conventional glass method, those target cells with a fluorescent signal-to-noise ratio of 12 could be microscopically observed at single-cell level within 2 to 5 min prior to secondary washing.