June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
A Rapid Multiplexed Ultrasensitive Nanoliter Lab-on-a-Chip Immunoassay Platform for Human Tear Film Analysis
Author Affiliations & Notes
  • Benjamin D Sullivan
    TearLab Corp, San Diego, CA
    Lµbris LLC, Boston, MA
  • Wendy Mendiola
    DCN, San Diego, CA
  • Melissa Lee
    TearLab Corp, San Diego, CA
  • Brandon Westerberg
    TearLab Corp, San Diego, CA
  • Matthew Solomon
    MiniFAB, Scoresby, VIC, Australia
  • Christian Potzner
    MiniFAB, Scoresby, VIC, Australia
  • Roy Chung
    DCN, San Diego, CA
  • Hans Boehringer
    DCN, San Diego, CA
  • Steve Zmina
    TearLab Corp, San Diego, CA
  • Erol Harvey
    MiniFAB, Scoresby, VIC, Australia
  • Footnotes
    Commercial Relationships Benjamin Sullivan, TearLab (E), TearLab (I), TearLab (P); Wendy Mendiola, TearLab (C); Melissa Lee, TearLab (E); Brandon Westerberg, TearLab (E); Matthew Solomon, TearLab (C); Christian Potzner, TearLab (C); Roy Chung, TearLab (C); Hans Boehringer, TearLab (C); Steve Zmina, TearLab (E), TearLab (I), TearLab (P); Erol Harvey, TearLab (C)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 322. doi:
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    • Get Citation

      Benjamin D Sullivan, Wendy Mendiola, Melissa Lee, Brandon Westerberg, Matthew Solomon, Christian Potzner, Roy Chung, Hans Boehringer, Steve Zmina, Erol Harvey; A Rapid Multiplexed Ultrasensitive Nanoliter Lab-on-a-Chip Immunoassay Platform for Human Tear Film Analysis. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):322.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract
 
Purpose
 

Existing methods for point of care molecular diagnostics require volumes of tear than are not readily available from dry eye disease patients. Manual sample transfers and long reaction times can compromise test integrity, reduce precision and impede clinical workflow. The purpose of this work was to categorize the dominant variables that influence nanoliter assay performance.

 
Methods
 

A three-dimensional microfluidic platform was constructed to accurately collect, meter and assay nanoliters of tear fluid. Integration of a long Stokes shifted fluorescent nanoparticle label with flow timing and homogenization structures facilitated an on-chip chromatographic platform for tear immunoassays. The geometry and chemistry of the upstream nanoparticle distribution was modulated to optimize the speed and sensitivity of the assay. Assays were run using 200 nL spiked human tear samples (BioreclamationIVT).

 
Results
 

After a 5 second incubation time, steady state signals were observed 20 seconds after co-localization of reactants, with a total sample-to-answer time under one minute. Test line integration from 45-60 seconds showed a significant difference (p < 2.4e-11) between control (1.59 ± 1.16 AU) vs. spiked samples (9.72 ± 0.95 AU), with a signal-to-noise ratio of 6.1 and a coefficient of variation of 9.7% for a 0.5 nM concentration (n=9 each). Significant differences in assay time and signal isotropy were observed for the various initial nanoparticle distributions.

 
Conclusions
 

Optimization of input signal distributions substantially improved sensitivity and speed of detection for point-of-care nanoliter assays.

 
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