Evanescent-imaging-ellipsometry-based microarray reader

Journal of Biomedical Optics
Srivatsa VenkatasubbaraoRussell Chipman

Abstract

We describe the development of a label-less ellipsometric imaging microarray reader. The ability of the ellipsometric microarray reader to measure binding of sample to microarray surface is verified using oligonucleotide complementary DNA (cDNA) microarrays. Polarized light illuminates the microarray surface through a glass substrate at an angle beyond the critical angle and changes in the polarization of totally internally reflected light resulting from binding events on the microarray surface are measured. This polarization change is used to measure the thickness of biomolecules bound to the microarray. A prototype ellipsometric imaging microarray reader is constructed and calibrated, and the performance is evaluated with cDNA microarrays. The microarray reader measures changes in refractive index changes as small as 0.0024 and thickness changes as small as 0.28 nm. The optimization of angle of incidence and substrate refractive index necessary to achieve high sensitivity is also described. This ellipsometric technique offers an attractive alternative to fluorescence-microarray readers in some genomic, proteomic, diagnostic, and sensing applications.

References

Nov 16, 2004·Trends in Biotechnology·Srivatsa Venkatasubbarao
Nov 1, 1992·Applied Optics·D H Goldstein

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Citations

May 28, 2008·Analytical and Bioanalytical Chemistry·Michael Seidel, Reinhard Niessner
Jun 15, 2007·Journal of Peptide Science : an Official Publication of the European Peptide Society·Vianney SoupletOleg Melnyk

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