High-throughput, dual probe biological assays based on single molecule detection

US7076092B2 · kind B2 · utility

Assignee

• United States Department of Energy · US

Inventors

• Christopher W. Hollars · Brentwood, US
• Thomas R. Huser · Werther, DE
• Stephen M. Lane · Oakland, US
• Rodney L. Balhorn · Livermore, US
• Olgica Bakajin · San Leandro, US
• Christopher B. Darrow · Pleasanton, US
• Joe H. Satcher, Jr. · Patterson, US

Key dates

• Filing date: Jun 12, 2002
• Grant date: Jul 11, 2006
• Expiry date: Jan 23, 2024

Classification

• Technology area (CPC G): Physics
• CPC primary: G01N2021/6441
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

A method and apparatus with the sensitivity to detect and identify single target molecules through the localization of dual, fluorescently labeled probe molecules. This can be accomplished through specific attachment of the taget to a surface or in a two-dimensional (2D) flowing fluid sheet having approximate dimensions of 0.5 μm×100 μm×100 μm. A device using these methods would have 103–104 greater throughput than previous one-dimensional (1D) micro-stream devices having 1 μm3 interrogation volumes and would for the first time allow immuno- and DNA assays at ultra-low (femtomolar) concentrations to be performed in short time periods (˜10 minutes). The use of novel labels (such as metal or semiconductor nanoparticles) may be incorporated to further extend the sensitivity possibly into the attomolar range.