A Double-Deck Self-Digitization Microfluidic Chip for Digital PCR

In this work, a double-deck microfluidic chip was presented for digital PCR application. This chip consists of two reverse-placed micro-patterned polydimethylsiloxane (PDMS) layers between the top and bottom glass substrates. Each micropatterned PDMS layer contains more than 20,000 cylindrical micro...

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Bibliographic Details
Published in:Micromachines (Basel) 2020-11, Vol.11 (12), p.1025
Main Authors: Xu, Gangwei, Si, Huaqing, Jing, Fengxiang, Sun, Peng, Zhao, Dan, Wu, Dongping
Format: Article
Language:English
Subjects:
Carbon black
Cylindrical chambers
Decks
Deoxyribonucleic acid
digital polymerase chain reaction
Digitization
Dilution
DNA
double-deck
Fluorescence
Genetic testing
Glass substrates
microfluidic chip
Microfluidics
Polydimethylsiloxane
Reagents
Silicon wafers
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Summary:In this work, a double-deck microfluidic chip was presented for digital PCR application. This chip consists of two reverse-placed micro-patterned polydimethylsiloxane (PDMS) layers between the top and bottom glass substrates. Each micropatterned PDMS layer contains more than 20,000 cylindrical micro-chambers to hold the partitioned droplets. The double-deck designs can double the number of chambers and reagent capacity without changing the planar area of the chip. In addition, carbon black was mixed into the pure PDMS gel to obstruct the passage of fluorescence from the positive chambers between the two layers of the chip. Thus, the fluorescence signal of micro-chambers in different layers of the chip after PCR can be collected without mutual interference. The quantitative capability of the proposed chip was evaluated by measuring a 10-fold serial dilution of the DNA template. A high accuracy of the absolute quantification for nucleic acid with a dynamic range of 10 was demonstrated by this chip in this work. Owing to its characteristics of small planar area, large capacity, and sensitivity, the double-deck microfluidic chip is expected to further promote the extensive applications of digital PCR.
ISSN:2072-666X
2072-666X
DOI:10.3390/mi11121025