Nanoreporter Coded Labeling for Single-Molecule Detection
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Solution Overview
Problem
Current microarray-based methods for detecting and quantifying target molecules in biomolecular samples are inefficient, requiring significant sample amounts and lacking the ability to detect molecules individually, with kinetics issues and limited sensitivity.
Innovation Solution
The development of coded, labeled reporter molecules (nanoreporters) that bind to target molecules, emitting unique light signals for identification and quantification, allowing for sensitive detection and analysis in small sample volumes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If microarray-based methods are used for detecting target molecules, then detection capability is provided, but significant sample amounts are required and individual molecule detection is not possible
Solution Approach 1:
The invention segments the detection target into individual molecules rather than measuring bulk concentration. Each nanoreporter binds to and detects individual target molecules, enabling single-molecule sensitivity. This segmentation allows detection of rare events and low-abundance targets without requiring microarray-scale sample amounts.
Solution Approach 2:
The invention uses coded nanoreporters as molecular copies that carry identification codes. These nanoreporters are designed to bind specifically to target molecules and emit coded signals, allowing detection of individual molecules through their unique codes rather than requiring bulk sample analysis.
2Productivity
If microarray-based methods are used, then detection is enabled, but kinetics are less efficient and quantification capability is limited
Solution Approach 1:
The invention replaces the mechanical surface-based hybridization system of microarrays with a solution-phase nanoreporter system. The nanoreporters perform hybridization in aqueous solution, which is kinetically faster and more efficient than surface-based binding. This substitution enables rapid detection without the time losses associated with microarray hybridization.
3Adaptability or versatility
If microarray technology is used, then simultaneous detection of multiple genes is possible, but individual molecule detection and direct quantification are not achievable
Solution Approach 1:
The invention applies local quality by assigning unique identification codes to individual nanoreporters. Each nanoreporter carries a specific code that allows its binding events to be tracked and quantified individually. This local identification enables precise measurement of individual molecule interactions while maintaining the ability to detect multiple target types simultaneously through code differentiation.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
Enables accurate and sensitive detection and quantification of individual target molecules, reducing sample requirements and improving analysis efficiency, facilitating diagnostic and prognostic applications.
Implementation Method 1
The formation of a duplex indicates the presence of the target sequence in the sample and the degree of duplex formation, as measured by the amount of label incorporated in it
Data Source
Figure 1A~1F
Figure 1D~1F
Figure 2A~2C
AI summary
Methods, computers, and computer program products for detecting the presence of a probe within a sample overlayed on a substrate are provided. The probe comprises a plurality of spatially arranged labels. A data storage module stores a plurality of light images, where each light image has light from the sample at a corresponding wavelength range in a plurality of different wavelength ranges. A label identification module identifies a plurality of labels in the plurality of light images that are proximate to each other on the substrate. A spatial order of the plurality of labels determines a string sequence of the plurality of labels. A probe identification module determines whether the string sequence of the plurality of labels comprises a valid reporter sequence.