PDMS-based opto-fluidic micro flow cytometer with two-color, multi-angle fluorescence detection capability using PIN photodiodes
PDMS-based opto-fluidic micro flow cytometer with two-color, multi-angle fluorescence detection capability using PIN photodiodes
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DOI：
10.1016/j.snb.2003.10.010
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发表时间：
2004-03-15
影响因子：
8.4
通讯作者：
Skerlos, SJ
中科院分区：
文献类型：
--
作者：
Tung, YC;Zhang, M;Skerlos, SJ
本研究描述了一种微流控观察通道的设计和性能，该通道能够使用固态激光器和基于pin的光电探测器进行流式细胞术测量。该观察单元采用基于聚二甲基硅氧烷（PDMS）的软光刻技术制造，设计有集成的微槽，允许光纤的物理注册，作为激光激发和荧光检测的光波导。光纤的排列设计允许在观察通道的单个询问点同时对样品粒子进行多色激发，使用多个激发和检测角度。多个角度可用于增加信噪比（SIN）通过执行两个独立的荧光测量同时进行，并允许多个同时的细胞术测量发生在一个单一的询问点。此外，多角度微流式细胞仪被证明可以提供一致的颜色区分，这使得该设计能够实现在传统流式细胞仪上不可能实现的光谱仪功能。通过检测荧光微球（15 μ m）和核酸标记真菌（Saccharomyces cerevisiae）来评价微流式细胞仪的性能。考察了荧光激发与检测之间的多个角度（45°、135°和180°）的S/N、S/N变异系数和颗粒回收率。实验证明了开发高信噪比单细胞荧光检测系统的可行性，该系统完全基于硅基PIN光电二极管和锁定放大技术。该技术适用于无机粒子和生物细胞发出的散射和荧光信号的检测，具有高度的经济化制造、小型化和低功耗操作的特点。因此，本研究中描述的设计方法和性能结果适用于开发实用和低成本的细胞计数微总量分析系统（muTAS）。(C) 2003 Elsevier B.V.版权所有
This research describes the design and performance of a microfluidic observation channel capable of performing flow cytometry measurements using solid-state lasers and PIN-based photodetectors. The observation cell, fabricated using poly(dimethylsiloxane) (PDMS)-based soft lithography, is designed with integrated microgrooves that permit the physical registration of optical fibers that serve as optical waveguides for laser excitation and fluorescence detection. The optical fiber arrangement is designed to permit simultaneous multi-color excitation of sample particles, at a single interrogation point in the observation channel, using multiple angles of excitation and detection. The multiple angles can be used to increase signal-to-noise ratio (SIN) by performing two independent fluorescence measurements simultaneously, and to permit multiple simultaneous cytometric measurements to occur at a single interrogation point. Moreover, the multi-angle micro flow cytometer is proven to provide consistent color differentiation that allows the design to achieve spectrometer capabilities not possible on a conventional flow cytometer. The micro flow cytometer performance was evaluated by considering the detection of fluorescent microspheres (15 mum) and nucleic acid labeled fungus (Saccharomyces cerevisiae). Multiple angles (45degrees, 135degrees, and 180degrees) between fluorescence excitation and detection were investigated with respect to S/N, S/N coefficient of variation, and particle recovery. The experiments demonstrate the feasibility of developing a high S/N single-cell fluorescence detection system based exclusively on silicon-based PIN photodiodes coupled with the lock-in amplification technique. The technique is applicable to the detection of scatter and fluorescent signals emanating from inorganic particles and biological cells, and is highly amenable to economical manufacturing, miniaturization, and low-power operation. Consequently, the design approach and performance results described in this investigation have applicability to the development of practical and low-cost cytometric micro total analysis systems (muTAS). (C) 2003 Elsevier B.V. All rights reserved.