Molecular imaging and related methods

Abstract

The present invention generally relates to imaging single molecules, or one or more collections of single molecules, and methods related to the imaging. In a method aspect, the present invention provides a method of imaging single molecules. The method comprises the steps of: a) exposing a test sample to a probe, wherein the probe comprises a first portion that specifically binds to a target molecule and a second portion that is detectable as the result of one or more chemical groups that interact with light at one or more wavelengths, wherein the probe binds to a target molecule to provide a complex; b) exposing the complex to one or more wavelengths of light that interact with the one or more chemical groups; c) detecting a result from the interacting of one or more wavelengths of light that interact with the one or more chemical groups to provide an image of one or more single molecules. The image possesses a resolution better than 450 nm over a view field area of at least 1×105 μm2, and wherein the image is obtained in a single detection step without variation of any detection settings.

Description

[0001] This application claims the benefit of the following US Provisional Patent Application Serial No. 61 / 851,276, filed March 6, 2013, which is incorporated herein by reference. technical field [0002] The present invention relates to the imaging of single molecules, or one or more collections of single molecules, and methods related to imaging. Background technique [0003] Methods have been reported to detect single molecules in extremely small areas (ie, areas smaller than 10 nm x 100 nm) with a resolution of about 300 nm. Fluorescence in situ hybridization (FISH), for example, is a method for measuring gene expression that is sensitive enough to detect individual mRNA molecules. As originally described by Singer, the method involves the simultaneous hybridization of five oligonucleotide probes to each mRNA target. Firmino AM, Fay FS, Fogarty K, Singh RH. Visualization of in situ transcription of single RNAs. "science". 1998;280:585-590. The oligonucleotides are ...

AI Technical Summary

Problems solved by technology

This method uses a large number of individually labeled probes instead of a limited number of multiplex labeled probes, and is used to overcome several problems caused by Singer's original FISH procedure: the large number of labeled oligonucleotides is difficult to synthesize and purify; when certain fluorescent Self-quenching occurs when groups are present in multiple copies in the same oligonucleotide; signal is prone to denaturation

Images