Assay Insert Reduces Autofluorescence in Detection
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Solution Overview
Problem
Current assay substrates face limitations in detection sensitivity due to autofluorescence and low binding capacity, particularly in fluorescence-based techniques, which interfere with accurate results.
Innovation Solution
The introduction of a system with a transparent window and an insert portion that reduces the air gap between the optical path and the substrate, combined with a polymer or membrane coating, enhances chemiluminescence detection and binding capacity by minimizing autofluorescence interference and optical aberrations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If nitrocellulose coating is applied to glass substrate, then binding capacity is improved, but autofluorescence interference increases
Solution Approach 1:
The patent introduces an insert portion as an intermediary component that physically separates the nitrocellulose coating from the glass substrate. This mediator allows the nitrocellulose to maintain its high binding capacity while preventing it from causing autofluorescence interference during detection, as the insert acts as a barrier between the binding surface and the detection optical path.
Solution Approach 2:
The patent segments the assay system into distinct functional components: the glass substrate provides structural support and optical transparency, while the insert portion containing the nitrocellulose coating provides the binding function. This segmentation allows each component to optimize its specific function without interfering with the other, particularly separating the binding function from the detection interference.
2Object-affected harmful factors
If plasma-treated plastic substrate is used, then autofluorescence interference is reduced, but binding capacity decreases
Solution Approach 1:
The patent merges the advantages of two different substrate materials by combining plasma-treated plastic (which provides low autofluorescence) with nitrocellulose coating (which provides high binding capacity). The insert portion structure allows the nitrocellulose to be applied on the plastic substrate, creating a composite system that exhibits both low autofluorescence and high binding capacity simultaneously.
3Device complexity
If air gap is present between optical path and substrate, then device simplicity is maintained, but optical aberrations increase
Solution Approach 1:
The insert portion acts as an intermediary optical element that fills or reduces the air gap between the detection optical path and the substrate. This mediator eliminates the fluid-air interface that causes surface tension effects and optical aberrations, while maintaining the overall simplicity of the device architecture.
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 improves the sensitivity of biochemical analyses by reducing autofluorescence interference and increasing binding capacity, allowing for more accurate chemiluminescence detection and reduced optical aberrations, thereby enhancing the overall effectiveness of assays.
Implementation Method 1
a nitrocellulose coating is applied to a surface of a glass plate, biological materials (e.g., proteins) are bound to the coating
Implementation Method 2
when light of a specific wavelength is shone upon the well, the antigen/antibody complexes will fluoresce
Implementation Method 3
a compound can be added to the well that causes the detect antibody to emit light within a predetermined wavelength (e.g., 400-500 nm)
Data Source
AI summary
Systems and methods are disclosed for improved detection sensitivity of assays. The systems include an upper plate that is inserted into a well of testing well, such as a base microtiter plate. For bottom detection, a coating, including polymer coatings or membrane coatings, is applied to an insert portion of the upper plate, and the base plate includes transparent windows. In bottom detection, a detector will image from below the base plate. Alternatively, for top detection, the coating is applied to the well of the base plate, and the upper plate includes transparent windows. In top detection, a detector will image from above the upper plate. Analysis features, including capture antibody features or antigen features, can be printed on the coating surface for forward-phase assays or reverse-phase assays, respectively. Methods for making and using the same are disclosed.