Highly efficient dual-channel cytometric-detection of micron-sized particles in microfluidic device

  1. Get@NRC: Highly efficient dual-channel cytometric-detection of micron-sized particles in microfluidic device (Opens in a new window)
DOIResolve DOI: http://doi.org/10.1016/j.snb.2010.09.055
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Journal titleSensors and Actuators, B: Chemical
Pages402409; # of pages: 8
SubjectAverage throughput; Cell count; Cytometric analysis; Detection accuracy; Detection efficiency; Dual channel; Dual channel detections; E. coli; Escherichia coli (E. coli); Fluorescein isothiocyanate; Fluorescence emission; Free-space optical; High-throughput measurements; Hydrodynamic focusing; Micro-fluidic devices; Microfabricated channels; Micron-sized particles; Mixing ratios; Non-pathogenic bacterium; Particle control; Polydimethylsiloxane PDMS; Scattering signals; Escherichia coli; Fluidic devices; Fluorescence; Light scattering; Microfabrication; Microfluidics; Refraction; Scattering; Silicones; Suspensions (fluids); Throughput; Signal detection
AbstractTo demonstrate the ability to efficiently count and identify suspended micron-sized particles by simultaneously detecting their fluorescence emission and light scattering in microfabricated channel, a compact configuration that used a polydimethylsiloxane (PDMS) microfabricated channel as interrogation component, hydrodynamic focusing for particle control, and a simple free-space optical setup for signal detection, was accordingly developed. Subsequently, a quantitative count of 1.013 μm diameter fluorescently labeled beads in suspension was implemented in a microfluidic device employing both fluorescence emission and light scattering at average particle throughput ranging from 83 to 416 particles/s. As a result, the detection efficiencies above 88% for both signals and correlation percentages above 97% between them were routinely achieved. In addition, it was shown that effective differentiation of 1.013 μm fluorescently labeled beads from various unlabeled beads in mixed populations of high mixing ratios had been successfully realized in this microfluidic-device-based instrumentation. Finally, the demonstrated system was used to detect fluorescein isothiocyanate (FITC) labeled nonpathogenic bacteria of Escherichia coli (E. coli) DH5α. The results showed the detection efficiencies above 89.7% for fluorescence emission and 94.5% for light scattering signals, and a correlation of 94.9% between the two signals at an average throughput of 350 cells/s have been obtained. As a comparison, the detection accuracies of the dual-channel cytometric detection of the FITC-labeled E. coli DH5α cells in the microfluidic device are approximately 84.3% and 88.8% for fluorescence emission and light scattering respectively when compared against a manual cell count using a haemocytometer as a standard. © 2010 Elsevier B.V. All rights reserved.
Publication date
AffiliationNational Research Council Canada (NRC-CNRC); NRC Institute for Microstructural Sciences (IMS-ISM)
Peer reviewedYes
NPARC number21271450
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Record identifier5e56e935-67a5-40c9-9625-c6c452b054c8
Record created2014-03-24
Record modified2016-05-09
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