A cell-based sensor of fluid shear stress for microfluidics.
A cell-based sensor of fluid shear stress for microfluidics.
复制标题
用于微流体的基于细胞的流体剪切应力传感器。
DOI：
10.1039/c4lc01369g
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发表时间：
2015
期刊：
影响因子：
6.1
通讯作者：
Voldman,Joel
中科院分区：
文献类型：
--
作者：
Varma,Sarvesh;Voldman,Joel
设计用于基于细胞的研究或应用的微系统固有地需要流体处理。在这样的系统内的流动不可避免地产生流体剪切应力（FSS），其可能不利地影响细胞健康。通常报告细胞活力、形态或生长的简单测定以指示对细胞生理学的任何严重干扰。然而，目前还没有直接的指标来专门评估微流控设备或竞争微流控技术中FSS的生理影响。本文介绍了第一个基因编码的细胞传感器，荧光定量的方式后，FSS途径激活。我们挑选了一个广泛使用的细胞系（NIH 3 T3 s），并创建了一个转录细胞传感器，当相关的FSS诱导蛋白质的转录开始时，荧光打开。具体来说，我们选择早期生长因子-1（一种机械敏感蛋白）上调作为FSS检测的节点。我们通过观察显微镜和流式细胞术观察到的响应于FSS途径的化学诱导的诱导荧光来验证我们的传感器途径特异性和功能性。重要的是，我们发现我们的细胞传感器可被一系列FSS强度和持续时间诱导，当应用30分钟时，检测极限为2达因cm-2。此外，我们的细胞传感器证明了它们的通用性，当使其流过具有典型流动条件的惯性微流体装置环境时显示出感应灵敏度。我们预计这些细胞传感器在微系统社区中有广泛的应用，允许设备设计师设计具有可接受的FSS的系统，并使最终用户能够评估FSS对他们感兴趣的测定的影响。
Microsystems designed for cell-based studies or applications inherently require fluid handling. Flows within such systems inevitably generate fluid shear stress (FSS) that may adversely affect cell health. Simple assays of cell viability, morphology or growth are typically reported to indicate any gross disturbances to cell physiology. However, no straightforward metric exists to specifically evaluate physiological implications of FSS within microfluidic devices, or among competing microfluidic technologies. This paper presents the first genetically encoded cell sensors that fluoresce in a quantitative fashion upon FSS pathway activation. We picked a widely used cell line (NIH3T3s) and created a transcriptional cell-sensor where fluorescence turns on when transcription of a relevant FSS-induced protein is initiated. Specifically, we chose Early Growth Factor-1 (a mechanosensitive protein) upregulation as the node for FSS detection. We verified our sensor pathway specificity and functionality by noting induced fluorescence in response to chemical induction of the FSS pathway, seen both through microscopy and flow cytometry. Importantly, we found our cell sensors to be inducible by a range of FSS intensities and durations, with a limit of detection of 2 dynes cm−2 when applied for 30 minutes. Additionally, our cell-sensors proved their versatility by showing induction sensitivity when made to flow through an inertial microfluidic device environment with typical flow conditions. We anticipate these cell sensors to have wide application in the microsystems community, allowing the device designer to engineer systems with acceptable FSS, and enabling the end-user to evaluate the impact of FSS upon their assay of interest.
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