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Oblique-incidence reflectivity difference microscope for label-free high-throughput detection of biochemical reactions in a microarray format.

Authors
  • X, Zhu
  • Jp, Landry
  • Ys, Sun
  • Jp, Gregg
  • Kit S. Lam
  • X, Guo
Type
Published Article
Journal
Applied Optics
Publisher
The Optical Society
Volume
46
Issue
10
Pages
1890–1895
Source
Kit Lam Lab
License
Unknown

Abstract

We describe a recently developed oblique-incidence reflectivity difference (OI-RD) microscope, a form of polarization-modulated imaging ellipsometer, for label-free-high-throughput detection of biomolecular reactions on DNA and protein microarrays. We present examples of application of this technique to end-point and real-time investigations of DNA-DNA hybridization, antibody-antigen capture, and protein-small-molecule binding reactions. Compared to a conventional imaging ellipsometer based on the polarizer-compensator-sample-analyzer scheme and under the off-null condition, a polarization-modulated OI-RD microscope is inherently more sensitive by at least 1 order of magnitude to thickness changes on a solid surface. Compared with imaging surface plasmon resonance microscopes based on reflectance change on falling or rising slopes of the surface plasmon resonance, the OI-RD microscope (1) has a comparable sensitivity, (2) is applicable to conventional microscope glass slides, and (3) easily covers a field of view as large as the entire surface of a 1 in. x 3 in. (2.54 cm x 7.62 cm) microscope slide.

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