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

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 .mu.m.times.100 .mu.m.times.100 .mu.m. A device using these methods would have 10.sup.3 10.sup.4 greater throughput than previous one-dimensional (1D) micro-stream devices having 1 .mu.m.sup.3 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 (.about.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.

Inventors:
 [1];  [2];  [3];  [2];  [4];  [5];  [6]
  1. Brentwood, CA
  2. Livermore, CA
  3. Oakland, CA
  4. San Leandro, CA
  5. Pleasanton, CA
  6. (Patterson, CA)
Issue Date:
Research Org.:
Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
908345
Patent Number(s):
7076092
Application Number:
10/170,876
Assignee:
The United States of America as represented by the United States Department of Energy (Washington, DC)
Patent Classifications (CPCs):
G - PHYSICS G01 - MEASURING G01N - INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
G - PHYSICS G06 - COMPUTING G06K - RECOGNITION OF DATA
DOE Contract Number:  
W-7405-ENG-48
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
47 OTHER INSTRUMENTATION

Citation Formats

Hollars, Christopher W, Huser, Thomas R, Lane, Stephen M, Balhorn, Rodney L, Bakajin, Olgica, Darrow, Christopher, and Satcher, Jr., Joe H. High-throughput, dual probe biological assays based on single molecule detection. United States: N. p., 2006. Web.
Hollars, Christopher W, Huser, Thomas R, Lane, Stephen M, Balhorn, Rodney L, Bakajin, Olgica, Darrow, Christopher, & Satcher, Jr., Joe H. High-throughput, dual probe biological assays based on single molecule detection. United States.
Hollars, Christopher W, Huser, Thomas R, Lane, Stephen M, Balhorn, Rodney L, Bakajin, Olgica, Darrow, Christopher, and Satcher, Jr., Joe H. Tue . "High-throughput, dual probe biological assays based on single molecule detection". United States. https://www.osti.gov/servlets/purl/908345.
@article{osti_908345,
title = {High-throughput, dual probe biological assays based on single molecule detection},
author = {Hollars, Christopher W and Huser, Thomas R and Lane, Stephen M and Balhorn, Rodney L and Bakajin, Olgica and Darrow, Christopher and Satcher, Jr., Joe H.},
abstractNote = {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 .mu.m.times.100 .mu.m.times.100 .mu.m. A device using these methods would have 10.sup.3 10.sup.4 greater throughput than previous one-dimensional (1D) micro-stream devices having 1 .mu.m.sup.3 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 (.about.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.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2006},
month = {7}
}

Patent:

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Works referenced in this record:

Single-molecule counting and identification in a microcapillary
journal, April 1998


Counting of Single Protein Molecules at Interfaces and Application of This Technique in Early-Stage Diagnosis
journal, August 1998


High-Throughput Single-Molecule Spectroscopy in Free Solution
journal, October 2000


Statistical Analysis of Single-Molecule Colocalization Assays
journal, March 2001


Confinement and Detection of Single Molecules in Submicrometer Channels
journal, August 1997