Composite Probe for Enzyme Metallographic Detection
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Solution Overview
Problem
Current biological detection methods face challenges in visualizing multiple targets simultaneously due to the limitations of black metallographic signals, which obscure overlapping stains, and the need for multiple labeling and detection procedures, compromising specimen integrity and resolution.
Innovation Solution
A composite probe comprising a directing agent, gold nanoparticles, and a redox active enzyme, which reacts with an enzyme substrate to produce a visible signal, allowing for simultaneous enzymatic and gold labeling in a single procedure, enabling enhanced sensitivity and correlation of information at different resolution levels.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If metallographic labeling is used for detection, then sensitivity and specificity are improved, but the black opaque signal obscures overlapping stains and prevents spectral imaging
Solution Approach 1:
The patent applies color changes by using gold nanoparticles that can be visualized at different wavelengths. The gold nanoparticles provide a spectral signal that can be detected across multiple wavelengths, allowing the system to distinguish between different targets based on their spectral signatures rather than relying on opaque black deposits. This enables spectral imaging while maintaining the high sensitivity of metallographic detection.
2Adaptability or versatility
If multiple labeling and detection procedures are used to detect multiple targets, then detection capability is improved, but specimen integrity is compromised and resolution is reduced
Solution Approach 1:
The patent merges multiple labeling and detection procedures into a single procedure by using composite probes that combine gold nanoparticles with enzymes or other detection molecules. This allows multiple targets to be detected simultaneously in one labeling step, maintaining specimen integrity and resolution while improving multi-target detection capability.
Solution Approach 2:
The patent applies universality by creating multi-functional probes that can detect multiple targets simultaneously. The gold nanoparticle-based probes can be designed to recognize different antigens or molecular targets, allowing a single probe system to perform multiple detection functions without requiring separate labeling procedures for each target.
3Measurement precision
If black metallographic signals are used for visualization, then detection sensitivity is improved, but overlapping targets cannot be resolved spectrally
Solution Approach 1:
The patent uses gold nanoparticles that exhibit characteristic spectral properties rather than producing opaque black signals. The gold nanoparticles can be detected across multiple wavelengths, creating a spectral signature that enables differentiation between overlapping targets based on their wavelength-specific signals, thus resolving the spectral resolution problem while maintaining detection sensitivity.
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
This approach facilitates the detection of multiple targets with increased sensitivity and resolution, allowing for correlative fluorescence or brightfield optical staining and electron microscopic labeling, without requiring multiple labeling operations, and provides higher detection sensitivity and improved visualization of biological targets.
Implementation Method 1
a redox active enzyme, for reacting with an enzyme substrate after the composite probe is bound to the target
Implementation Method 2
at least one nanoparticle conjugated to the at least one directing agent, the at least one nanoparticle comprising a plurality of gold atoms
Data Source
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Figure 3a~3d
Figure 4a~4b
AI summary
A test agent includes a composite probe having at least one nanoparticle having multiple metal atoms, a directing agent, and an enzyme. The directing agent attaches the probe to a target in a test sample. The test sample and bound probe are then treated with an enzyme substrate. A method of detecting a target in a test sample includes exposing the test sample to the probe, then treating the test sample with an enhancement or development solution to deposit at least one of a fluorophore, a chromgen, or a metal.