Spatially selective photonic crystal enhanced fluorescence and application to background reduction for biomolecule detection assays

Vikram Chaudhery, Cheng Sheng Huang, Anusha Pokhriya, James Polans, Brian T. Cunningham

Research output: Contribution to journalArticlepeer-review

Abstract

By combining photonic crystal label-free biosensor imaging with photonic crystal enhanced fluorescence, it is possible to selectively enhance the fluorescence emission from regions of the PC surface based upon the density of immobilized capture molecules. A label-free image of the capture molecules enables determination of optimal coupling conditions of the laser used for fluorescence imaging of the photonic crystal surface on a pixel-bypixel basis, allowing maximization of fluorescence enhancement factor from regions incorporating a biomolecule capture spot and minimization of background autofluorescence from areas between capture spots. This capability significantly improves the contrast of enhanced fluorescent images, and when applied to an antibody protein microarray, provides a substantial advantage over conventional fluorescence microscopy. Using the new approach, we demonstrate detection limits as low as 0.97 pg/ml for a representative protein biomarker in buffer.

Original languageEnglish (US)
Pages (from-to)23327-23340
Number of pages14
JournalOptics Express
Volume19
Issue number23
DOIs
StatePublished - Nov 7 2011

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics

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