A hard microflow cytometer using groove-generated sheath flow for multiplexed bead and cell assays |
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Authors: | Abel L Thangawng Jason S Kim Joel P Golden George P Anderson Kelly L Robertson Vyechi Low Frances S Ligler |
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Institution: | (1) Center for Bio/Molecular Science and Engineering, Naval Research Laboratory, Washington, DC 20375-5348, USA; |
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Abstract: | With a view toward developing a rugged microflow cytometer, a sheath flow system was micromachined in hard plastic (polymethylmethacrylate)
for analysis of particles and cells using optical detection. Six optical fibers were incorporated into the interrogation region
of the chip, in which hydrodynamic focusing narrowed the core stream to ∼35 μm × 40 μm. The use of a relatively large channel
at the inlet as well as in the interrogation region (375 μm × 125 μm) successfully minimized the risk of clogging. The device
could withstand pressures greater than 100 psi without leaking. Assays using both coded microparticles and cells were demonstrated
using the microflow cytometer. Multiplexed immunoassays detected nine different bacteria and toxins using a single mixture
of coded microspheres. A549 cancer cells processed with locked nucleic acid probes were evaluated using fluorescence in situ
hybridization. |
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