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TIRF-enhanced nanobeads’ Brownian diffusion measurements for detecting CRP in human serum

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Abstract

This paper reports the in situ detection of C-reactive protein (CRP) and antibody-coated nanobead interaction by measuring the variation of the Brownian diffusion of the nanobeads. The measurements are carried out by using total internal reflective fluorescence-enhanced (TIRF-enhanced) micro-particle-tracking velocimetry (micro-PTV), which is highly sensitive for analyzing nanobead movements in the near-wall region. The characteristics of evanescent wave penetration of various liquid samples, which have different refractive indices, are investigated for nanobead illumination. The Brownian velocities of the nanobeads in various concentrations of CRPs within glycerol solution and serum are measured in real time. The detectable CRP concentration range of 1–6 mg/ml in human serum is found, and the detecting process can be finished in 10 min. From the results, the dissociation constant of CRP and anti-CRP in various viscosity solutions can be determined through the developed kinetic analysis.

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Acknowledgments

This work was supported by National Science Council of Taiwan under grant numbers NSC 102-2627-E-002-002 and NSC 100-2221-E-002-107-MY3.

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Authors and Affiliations

  1. Institute of Applied Mechanics, National Taiwan University, No. 1, Section 4, Roosevelt Road, Taipei, 106, Taiwan, ROC

    Yu-Jui Fan, Horn-Jiunn Sheen, Zheng-Yu Chen & Kuang-Chong Wu

  2. Engineering Science and Ocean Engineering, National Taiwan University, No.1, Section 4, Roosevelt Road, Taipei, 106, Taiwan, ROC

    Yi-Hsing Liu & Jing-Fa Tsai

Authors
  1. Yu-Jui Fan
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  2. Horn-Jiunn Sheen
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  3. Zheng-Yu Chen
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  4. Yi-Hsing Liu
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  5. Jing-Fa Tsai
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  6. Kuang-Chong Wu
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Corresponding author

Correspondence to Horn-Jiunn Sheen.

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Fan, YJ., Sheen, HJ., Chen, ZY. et al. TIRF-enhanced nanobeads’ Brownian diffusion measurements for detecting CRP in human serum. Microfluid Nanofluid 19, 85–94 (2015). https://doi.org/10.1007/s10404-015-1551-y

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  • DOI: https://doi.org/10.1007/s10404-015-1551-y

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