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Issue 22, 2011
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1-Million droplet array with wide-field fluorescence imaging for digital PCR

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Abstract

Digital droplet reactors are useful as chemical and biological containers to discretize reagents into picolitre or nanolitre volumes for analysis of single cells, organisms, or molecules. However, most DNA based assays require processing of samples on the order of tens of microlitres and contain as few as one to as many as millions of fragments to be detected. Presented in this work is a droplet microfluidic platform and fluorescence imaging setup designed to better meet the needs of the high-throughput and high-dynamic-range by integrating multiple high-throughput droplet processing schemes on the chip. The design is capable of generating over 1-million, monodisperse, 50 picolitre droplets in 2–7 minutes that then self-assemble into high density 3-dimensional sphere packing configurations in a large viewing chamber for visualization and analysis. This device then undergoes on-chip polymerase chain reaction (PCR) amplification and fluorescence detection to digitally quantify the sample's nucleic acid contents. Wide-field fluorescence images are captured using a low cost 21-megapixel digital camera and macro-lens with an 8–12 cm2 field-of-view at 1× to 0.85× magnification, respectively. We demonstrate both end-point and real-time imaging ability to perform on-chip quantitative digital PCR analysis of the entire droplet array. Compared to previous work, this highly integrated design yields a 100-fold increase in the number of on-chip digitized reactors with simultaneous fluorescence imaging for digital PCR based assays.

Graphical abstract: 1-Million droplet array with wide-field fluorescence imaging for digital PCR

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Supplementary files

Publication details

The article was received on 27 Jun 2011, accepted on 25 Aug 2011 and first published on 29 Sep 2011


Article type: Paper
DOI: 10.1039/C1LC20561G
Citation: Lab Chip, 2011,11, 3838-3845
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    1-Million droplet array with wide-field fluorescence imaging for digital PCR

    A. C. Hatch, J. S. Fisher, A. R. Tovar, A. T. Hsieh, R. Lin, S. L. Pentoney, D. L. Yang and A. P. Lee, Lab Chip, 2011, 11, 3838
    DOI: 10.1039/C1LC20561G

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