Detection of biomolecular microarrays without fluorescent-labeling agents

J. P. Landry, X. D. Zhu, Jeffrey Gregg, X. W. Guo

Research output: Chapter in Book/Report/Conference proceedingConference contribution

11 Scopus citations

Abstract

We developed an optical oblique-incidence reflectivity difference (OI-RD) scanning microscope for imaging microarrays of label-free protein and DNA. Such a microscope complements currently widely used fluorescence-based optical microscopes by offering the capability to detect biochemical activities of DNA and protein molecules without the influence of fluorescent-labeling molecules. The specific activity and function of protein molecules are particularly subject to binding of small or large foreign molecules either directly through conformational change in the protein molecule itself or indirectly through properties of the attached molecules. We show that an OI-RD microscope can be used to: 1) detect binding reactions on microarrays without labeling, 2) quantitatively measure the optical properties of microarray spots, and 3) detect microarrays submerged in solution (which will enable OI-RD to monitor reaction kinetics on microarrays in reactive solutions). Furthermore, using both OI-RD and fluorescence images of an immunoglobulin-G (IgG) protein microarray, we observed that labeled and unlabeled IgG molecules deposited on the microarray substrate exhibit different wetting behaviors, and a mixture of the two tends to segregate into labeled and unlabeled regions. This illustrates potentially undesirable effects of fluorescent-labeling agents on protein properties that are of interest.

Original languageEnglish (US)
Title of host publicationProceedings of SPIE - The International Society for Optical Engineering
EditorsD.V. Nicolau, R. Raghavachari
Pages121-128
Number of pages8
Volume5328
DOIs
StatePublished - 2004
EventMicroarrays and Combinatorial Techniques: Design, Fabrication, and Analysis II - San Jose, CA, United States
Duration: Jan 25 2004Jan 26 2004

Other

OtherMicroarrays and Combinatorial Techniques: Design, Fabrication, and Analysis II
CountryUnited States
CitySan Jose, CA
Period1/25/041/26/04

Keywords

  • Biochips
  • Ellipsometry
  • Hybridization
  • Immunoglobulin-G
  • In-situ detection
  • Label-free detection
  • Microarrays
  • Oblique-incidence reflectivity difference
  • OI-RD
  • Oligonucleotide

ASJC Scopus subject areas

  • Electrical and Electronic Engineering
  • Condensed Matter Physics

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  • Cite this

    Landry, J. P., Zhu, X. D., Gregg, J., & Guo, X. W. (2004). Detection of biomolecular microarrays without fluorescent-labeling agents. In D. V. Nicolau, & R. Raghavachari (Eds.), Proceedings of SPIE - The International Society for Optical Engineering (Vol. 5328, pp. 121-128) https://doi.org/10.1117/12.524554