Molecular Imaging and Related Methods

Abstract

The present invention relates to the imaging of single molecules, or one or more collections of single molecules, and methods related to imaging. In terms of methods, the present invention provides methods for single molecule imaging. The method comprises the steps of: a) exposing a test sample to a probe, wherein the probe comprises a first moiety that specifically binds to a target molecule and one or more chemical moieties that interact with light at one or more wavelengths. group and detectable second moiety, wherein the probe binds to the target molecule to form a complex; b) exposing the complex to one or more of the light interacting with the one or more chemical groups wavelength, and; c) light interacting with said one or more chemical groups, detecting the result of the interaction with said one or more wavelengths, generating one or more single-molecule images. The image has a resolution higher than 450 nanometers over an imaging area of ​​at least 1 x 105 square microns, and wherein the image is obtained in a single detection step without changing 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