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Integrated imaging instrument for self-calibrated fluorescence protein microarrays

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3 Author(s)
Reddington, A.P. ; Department of Electrical and Computer Engineering, Boston University, Boston, Massachusetts 02215, USA ; Monroe, M.R. ; Unlu, M.S.

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Protein microarrays, or multiplexed and high-throughput assays, monitor multiple protein binding events to facilitate the understanding of disease progression and cell physiology. Fluorescence imaging is a popular method to detect proteins captured by immobilized probes with high sensitivity and specificity. Reliability of fluorescence assays depends on achieving minimal inter- and intra-assay probe immobilization variation, an ongoing challenge for protein microarrays. Therefore, it is desirable to establish a label-free method to quantify the probe density prior to target incubation to calibrate the fluorescence readout. Previously, a silicon oxide on silicon chip design was introduced to enhance the fluorescence signal and enable interferometric imaging to self-calibrate the signal with the immobilized probe density. In this paper, an integrated interferometric reflectance imaging sensor and wide-field fluorescence instrument is introduced for sensitive and calibrated microarray measurements. This platform is able to analyze a 2.5 mm × 3.4 mm area, or 200 spots (100 μm diameter with 200 μm pitch), in a single field-of-view.

Published in:

Review of Scientific Instruments  (Volume:84 ,  Issue: 10 )

Date of Publication:

Oct 2013

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