TY - JOUR
T1 - High sensitivity automated multiplexed immunoassays using photonic crystal enhanced fluorescence microfluidic system
AU - Tan, Yafang
AU - Tang, Tiantian
AU - Xu, Haisheng
AU - Zhu, Chenqi
AU - Cunningham, Brian T.
N1 - Funding Information:
This work was supported by Grants from the National Institutes of Health ( GM086382A , R33CA177446 , R01GM108584 ) and the National Science Foundation (CBET 07-54122 ). Any opinions, findings, conclusions, or recommendations expressed in this material are those of the authors and do not necessarily reflect the views of the National Institutes of Health or the National Science Foundation. The authors thank Scott McDonald and David Switzer at machine shop of ECE department for their help making the holding fixture.
Publisher Copyright:
© 2015 .
PY - 2015/11/5
Y1 - 2015/11/5
N2 - We demonstrate a platform that integrates photonic crystal enhanced fluorescence (PCEF) detection of a surface-based microspot fluorescent assay with a microfluidic cartridge to achieve simultaneous goals of high analytic sensitivity (single digit pg/mL), high selectivity, low sample volume, and assay automation. The PC surface, designed to provide optical resonances for the excitation wavelength and emission wavelength of Cyanines 5 (Cy5), was used to amplify the fluorescence signal intensity measured from a multiplexed biomarker microarray. The assay system is comprised of a plastic microfluidic cartridge for holding the PC and an assay automation system that provides a leak-free fluid interface during introduction of a sequence of fluids under computer control. Through the use of the assay automation system and the PC embedded within the microfluidic cartridge, we demonstrate pg/mL-level limits of detection by performing representative biomarker assays for interleukin 3 (IL3) and Tumor Necrosis Factor (TNF-α). The results are consistent with limits of detection achieved without the use of the microfluidic device with the exception that coefficients of variability from spot-to-spot are substantially lower than those obtained by performing assays with manual manipulation of assay liquids. The system's capabilities are compatible with the goal of diagnostic instruments for point-of-care settings.
AB - We demonstrate a platform that integrates photonic crystal enhanced fluorescence (PCEF) detection of a surface-based microspot fluorescent assay with a microfluidic cartridge to achieve simultaneous goals of high analytic sensitivity (single digit pg/mL), high selectivity, low sample volume, and assay automation. The PC surface, designed to provide optical resonances for the excitation wavelength and emission wavelength of Cyanines 5 (Cy5), was used to amplify the fluorescence signal intensity measured from a multiplexed biomarker microarray. The assay system is comprised of a plastic microfluidic cartridge for holding the PC and an assay automation system that provides a leak-free fluid interface during introduction of a sequence of fluids under computer control. Through the use of the assay automation system and the PC embedded within the microfluidic cartridge, we demonstrate pg/mL-level limits of detection by performing representative biomarker assays for interleukin 3 (IL3) and Tumor Necrosis Factor (TNF-α). The results are consistent with limits of detection achieved without the use of the microfluidic device with the exception that coefficients of variability from spot-to-spot are substantially lower than those obtained by performing assays with manual manipulation of assay liquids. The system's capabilities are compatible with the goal of diagnostic instruments for point-of-care settings.
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U2 - 10.1016/j.bios.2015.05.041
DO - 10.1016/j.bios.2015.05.041
M3 - Article
C2 - 26043313
AN - SCOPUS:84930633575
VL - 73
SP - 32
EP - 40
JO - Biosensors and Bioelectronics
JF - Biosensors and Bioelectronics
SN - 0956-5663
ER -