Oil-Based Microfluidic Chip for NASBA Reactions

The SD chip was fabricated as described previously.^{27 (link)} An oil mixture composed of 0.04% Abil WE 09, 91% Tegosoft DEC, and 9% light mineral oil was prepared and pipetted into the inlet and outlet of the chip. A vacuum was used to load the oil onto the chip, displacing air from the channel and chamber array. The NASBA reaction mixture was added to the inlet, and flowed into the channel and chambers under vacuum. An additional oil mixture was then added to isolate the aqueous sample in compartmentalized volumes, digitizing the sample. For digitization tests, 2 μM calcein was added to the NASBA buffer to generate a fluorescent signal. Fluorescent photographs were acquired with an Olympus MVX10 stereoscope (Olympus Corporation, Shinjuku, Tokyo, Japan).

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Wang J., Kreutz J.E., Thompson A.M., Qin Y., Sheen A.M., Wang J., Wu L., Xu S., Chang M., Raugi D.N., Smith R.A., Gottlieb G.S, & Chiu D.T. (2018). SD-Chip Enabled Quantitative Detection of HIV RNA using Digital Nucleic Acid Sequence-Based Amplification (dNASBA). Lab on a chip, 18(22), 3501-3506.

Publication 2018

MVX10 stereoscope

Buffer Calcein Chip Light Mineral oil Nasba Vacuum

Corresponding Organization : Seattle University

Other organizations : Fred Hutch Cancer Center, National Institute of Allergy and Infectious Diseases

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Variable analysis

independent variables

Fabrication of the SD chip as described previously
Preparation of the oil mixture composed of 0.04% Abil WE 09, 91% Tegosoft DEC, and 9% light mineral oil
Addition of 2 μM calcein to the NASBA buffer

dependent variables

Displacement of air from the channel and chamber array by the oil mixture
Flow of the NASBA reaction mixture into the channel and chambers under vacuum
Isolation of the aqueous sample in compartmentalized volumes
Generation of a fluorescent signal

control variables

Pipetting the oil mixture into the inlet and outlet of the chip
Use of a vacuum to load the oil onto the chip
Addition of the NASBA reaction mixture to the inlet

positive control

Not specified

negative control

Not specified

Annotations

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