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使用手机摄像头和时域编码光流控技术实现超高通量检测(每秒 100 万个荧光液滴)。

Ultra-high throughput detection (1 million droplets per second) of fluorescent droplets using a cell phone camera and time domain encoded optofluidics.

机构信息

Department of Bioengineering, School of Engineering and Applied Sciences, University of Pennsylvania, Philadelphia, Pennsylvania, USA.

Department of Chemical and Biomolecular Engineering, University of Pennsylvania, Philadelphia, Pennsylvania, USA.

出版信息

Lab Chip. 2017 Mar 14;17(6):1083-1094. doi: 10.1039/c6lc01489e.

Abstract

Droplet-based assays-in which ultra-sensitive molecular measurements are made by performing millions of parallel experiments in picoliter droplets-have generated enormous enthusiasm due to their single molecule resolution and robustness to reaction conditions. These assays have great untapped potential for point of care diagnostics but are currently confined to laboratory settings due to the instrumentation necessary to serially generate, control, and measure tens of millions of droplets. To address this challenge, we have developed the microdroplet megascale detector (μMD) that can generate and detect the fluorescence of millions of droplets per second (1000× faster than conventional approaches) using only a conventional cell phone camera. The key innovation of our approach is borrowed from the telecommunications industry, wherein we modulate the excitation light with a pseudorandom sequence that enables individual droplets to be resolved that would otherwise overlap due to the limited frame rate of digital cameras. Using this approach, the μMD measures droplets at a rate of 10 droplets per sec (ϕ = 166 mL h) in 120 parallel microfluidic channels and achieves a limit of detection LOD = 1 μM Rhodamine dye, sufficient for typical droplet based assays. We incorporate this new droplet detection technology with our previously reported parallelized droplet production strategy, incorporating 200 parallel droplet makers and only one set of continuous and droplet phase inputs and one output line. By miniaturizing and integrating droplet based diagnostics into a handheld format, the μMD platform can translate ultra-sensitive droplet based assays into a self-contained platform for practical use in clinical and industrial settings.

摘要

基于液滴的分析方法——通过在皮升级别的液滴中进行数百万个平行实验来进行超灵敏的分子测量——由于其单分子分辨率和对反应条件的稳健性,引起了极大的兴趣。这些分析方法在即时诊断方面具有巨大的未开发潜力,但由于需要连续生成、控制和测量数千万个液滴的仪器,目前仅限于实验室环境。为了解决这个挑战,我们开发了微滴兆级检测器 (μMD),它可以使用普通的手机摄像头每秒生成和检测数百万个液滴的荧光(比传统方法快 1000 倍)。我们方法的关键创新是借鉴电信行业的方法,我们用伪随机序列调制激发光,从而可以分辨单个液滴,否则由于数字相机的有限帧率,这些液滴会因重叠而无法分辨。使用这种方法,μMD 在 120 个平行微流道中以 10 个液滴/秒(ϕ = 166 mL h)的速度测量液滴,并实现检测限 LOD = 1 μM 若丹明染料,足以满足典型的基于液滴的分析。我们将这种新的液滴检测技术与我们之前报道的并行化液滴生成策略相结合,该策略整合了 200 个并行液滴生成器,仅使用一组连续和液滴相输入和一个输出线。通过将基于液滴的诊断微型化和集成到手持式格式中,μMD 平台可以将超灵敏的基于液滴的分析转化为一个独立的平台,以便在临床和工业环境中实际使用。

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