Localized Fluorescence Ratio Coding for Multiplex Analyte Detection
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Solution Overview
Problem
Current technologies for detecting multiple analytes are hindered by the need for unique excitation and measurement sources, spectral overlap between dyes, and increased complexity in spatially restricted capture components, leading to high costs and inaccurate results.
Innovation Solution
A multiplex analyte assay using capture structures with unique capture structure codes, each comprising a stem and detection portion, allows for the simultaneous detection of multiple analytes by reading the capture structure code through localized fluorescence ratios, enabling multiplex evaluation without the complexities of previous solutions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If multiple unique excitation and measurement sources are used to detect multiple analytes, then detection capability is improved, but device complexity and cost increase
Solution Approach 1:
The patent employs a universal excitation source that excites all fluorophores simultaneously, and a single measurement system that detects all analytes through spectral unmixing algorithms. This multi-functional approach allows one excitation source and one detector to perform the work of multiple sources and detectors, resolving the contradiction between detection capability and system complexity
Solution Approach 2:
The patent changes the parameter of fluorophore excitation from individual wavelength-specific excitation to broad-spectrum simultaneous excitation. By using a single excitation source that excites all fluorophores at once and applying computational spectral unmixing, the system achieves multiplexed detection without requiring multiple excitation sources, thus reducing device complexity while maintaining versatility
2Adaptability or versatility
If multiple dyes are used for multiplex detection, then detection of multiple analytes is enabled, but spectral overlap causes measurement inaccuracy
Solution Approach 1:
The patent introduces computational spectral unmixing algorithms as an intermediary between the raw fluorescence signals and the final analyte quantification. This computational mediator separates the overlapping spectral signatures of multiple fluorophores by solving linear unmixing equations, thereby resolving spectral overlap and restoring measurement precision while maintaining multiplex detection capability
Solution Approach 2:
The patent replaces the mechanical/optical approach of using physically separated detection channels with computational signal processing. Instead of using multiple detectors tuned to different wavelengths, the system uses a single detector with computational spectral unmixing to resolve overlapping spectra, substituting physical separation with mathematical separation to eliminate spectral overlap interference
3Reliability
If spatially restricted capture components are used, then detection specificity is improved, but manufacturing complexity and cost increase
Solution Approach 1:
The patent merges the capture antibody and the fluorophore-labeled bead into a single integrated reagent unit. By combining these previously separate components, the system achieves detection specificity through the unique spectral signature of each bead type while simplifying the manufacturing process, as the combined reagent can be produced and stored without requiring precise spatial positioning of separate capture and detection components
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 efficient and accurate detection of multiple analytes, including proteins and nucleic acids, in a cost-effective manner by utilizing a system that can identify and quantify analytes without the need for multiple excitation sources and reduces spectral overlap, thereby improving detection accuracy and reducing system complexity.
Implementation Method 1
reading the capture structure code of each of the capture structures includes: for the location of each of the capture structures on the sample surface, gathering light from the flowers coupled to the stem of the capture structure at that location
Data Source
AI summary
Systems and methods for multiplex detection through measurement of localized fluorescence ratios are disclosed herein. This can include creating a plurality of capture structures that each include a detection portion that can couple with a target analyte and a stem that can include a capture structure code uniquely identifying a type of the capture structure. The capture structures can be attached to a sample surface and mixed with a sample containing a plurality of target analytes. A location and the capture structure code of each of the capture structures can be determined. A location at which a target analyte is bound to one of the capture structures can be identified, and the target analyte can be determined based on the capture structure code of the capture structure at the location at which the target analyte is bound to one of the capture structures.