Out-of-plane integration of a multimode optical fiber for single particle/cell detection at multiple points on a microfluidic device with applications to particle/cell counting, velocimetry, size discrimination and the analysis of single cell lysate injec
Out-of-plane integration of a multimode optical fiber for single particle/cell detection at multiple points on a microfluidic device with applications to particle/cell counting, velocimetry, size discrimination and the analysis of single cell lysate injec
复制标题
多模光纤的面外集成，用于微流体装置上多个点的单颗粒/细胞检测，并应用于颗粒/细胞计数、测速、尺寸辨别和单细胞裂解物注射分析
DOI：
10.1039/c6lc01161f
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发表时间：
2017
期刊：
影响因子：
6.1
通讯作者：
Culbertson, Christopher T.
中科院分区：
文献类型：
--
作者：
Sadeghi, Jalal;Patabadige, Damith E.;Culbertson, Anne H.;Latifi, Hamid;Culbertson, Christopher T.
本文报道了一种集成面外多模光纤（OP-MMF）的单粒子/细胞跟踪微流控装置。该OP-MMF仅使用一个激发源和一个检测器就可以产生三条激发光线和三个检测点。它利用光学隧道模式在从单个光纤端发出的微流体通道中产生两条激发线。该方法用于准确计数颗粒/细胞，并进行速度测量和尺寸区分。使用OP-MMF测定了5、7和10 μm荧光标记的聚苯乙烯珠的速度和尺寸分布。此外，该方法还用于利用分离通道中的第三激发线分析细胞裂解物。OP-MMF装置在裂解前准确检测完整细胞两次，测定其速度，并在分离通道中进样点下游3 mm处检测进样的细胞裂解物。使用该设置，以自动化方式确定细胞进入裂解交叉点的速度和注入分离通道的荧光标记分析物的绝对迁移时间。该方法使我们能够使用来自注射的裂解物信号和来自裂解前的完整细胞的信号来确定裂解/注射效率系数（K）。K提供了注射到分离通道中的细胞裂解物的量的可靠测量。这里报道的方法可以在未来用于跟踪各种现有微流体装置中的颗粒、细胞或液滴，而不需要多路复用的掩模、层、庞大的光学元件或复杂的光学设计。
In this paper a single particle/cell-tracking microfluidic device that integrates an out-of-plane multimode optical fiber (OP-MMF) is reported. This OP-MMF is used to generate three excitation light-lines and three detection spots using only one excitation source and one detector. It takes advantage of an optical tunneling mode to create two excitation lines in a microfluidic channel emanating from a single fiber end. This method was used to accurately count particles/cells and perform velocity measurements and size discrimination. The velocity and size distributions of 5, 7, and 10 μm fluorescently labeled polystyrene beads were determined using the OP-MMF. Additionally, this method was used to analyze cell lysates with the third excitation line in the separation channel. The OP-MMF setup accurately detected an intact cell twice ∼2 mm prior to lysis, determined its velocity, and detected the injected cell lysate 3 mm downstream of the injection point in the separation channel. Using this setup, the velocity of cells entering the lysis intersection and the absolute migration times of fluorescently labeled analytes injected into the separation channel were determined in an automated fashion. This method enabled us to determine a lysing/injection efficiency coefficient (K) using signals from the injected lysate signal and from the intact cell before lysing. K provided a reliable measurement of the amount of cell lysate that was injected into the separation channel. The approach reported here could be used in the future to track particles, cells or droplets in a variety of existing microfluidic devices without the need for multiplexed masks, layers, bulky optical elements or complex optical designs.