title: Integrated imaging instrument for self-calibrated fluorescence protein microarrays.
abstract: 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.
authors.affiliation: Department of Electrical and Computer Engineering, Boston University, Boston, Massachusetts 02215, USA.
authors.forename: A P
authors.initials: AP
authors.lastname: Reddington
authors.forename: M R
authors.initials: MR
authors.lastname: Monroe
authors.forename: M S
authors.initials: MS
authors.lastname: Ünlü
mesh_terms: D000818:Animals; D002138:Calibration; D002417:Cattle; D004867:Equipment Design; D007368:Interferometry; D061848:Optical Imaging / Q000295:instrumentation / Q000379:methods*; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods*; D011817:Rabbits; D012984:Software