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Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 1, Iss. 3 — Oct. 1, 2010
  • pp: 983–997

Mass-transport limitations in spot-based microarrays

Ming Zhao, Xuefeng Wang, and David Nolte  »View Author Affiliations

Biomedical Optics Express, Vol. 1, Issue 3, pp. 983-997 (2010)

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Mass transport of analyte to surface-immobilized affinity reagents is the fundamental bottleneck for sensitive detection in solid-support microarrays and biosensors. Analyte depletion in the volume adjacent to the sensor causes deviation from ideal association, significantly slows down reaction kinetics, and causes inhomogeneous binding across the sensor surface. In this paper we use high-resolution molecular interferometric imaging (MI2), a label-free optical interferometry technique for direct detection of molecular films, to study the inhomogeneous distribution of intra-spot binding across 100 micron-diameter protein spots. By measuring intra-spot binding inhomogeneity, reaction kinetics can be determined accurately when combined with a numerical three-dimensional finite element model. To ensure homogeneous binding across a spot, a critical flow rate is identified in terms of the association rate ka and the spot diameter. The binding inhomogeneity across a spot can be used to distinguish high-affinity low-concentration specific reactions from low-affinity high-concentration non-specific binding of background proteins.

© 2010 OSA

ToC Category:
Biosensors and Molecular Diagnostics

Original Manuscript: August 2, 2010
Revised Manuscript: September 16, 2010
Manuscript Accepted: September 17, 2010
Published: September 20, 2010

Ming Zhao, Xuefeng Wang, and David Nolte, "Mass-transport limitations in spot-based microarrays," Biomed. Opt. Express 1, 983-997 (2010)

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