Microparticle Detection via Bright and Dark Field Imaging
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Solution Overview
Problem
Current biomolecule detection methods face challenges such as complex processes, low stability, low sensitivity, low accuracy, and high cost, which hinder their clinical application and efficiency in detecting low-abundance molecules like neurological factors and cancer factors.
Innovation Solution
The method involves using a common solid phase support like a glass slide, multi-well plate, or flow channel for random uniform distribution of microparticles, followed by direct imaging under bright and dark fields, simplifying the detection process and enhancing sensitivity, accuracy, and cost-effectiveness.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If chemiluminescent immunoassay is used for detection, then the detection process can be performed with existing technology, but the detection sensitivity and dynamic range are limited
Solution Approach 1:
The invention divides the detection system into two independent counting channels: bright field counting for total microparticle concentration and dark field counting for signal molecule-containing microparticles. This segmentation allows digital quantification of low-abundance molecules by calculating the ratio of dark field to bright field counts, thereby achieving high detection sensitivity without increasing overall system complexity
Solution Approach 2:
The invention replaces the chemical luminescence reaction mechanism with direct optical imaging and digital counting. Instead of relying on chemiluminescent reactions that have inherent sensitivity limits, the system uses bright field and dark field microscopy to directly visualize and count microparticles, substituting chemical detection with optical detection mechanisms
2Measurement precision
If digital immunodiagnosis technology is implemented, then detection sensitivity can be improved, but the detection process becomes more complex
Solution Approach 1:
The invention uses a universal microparticle platform that can be applied to detect various signal molecules (proteins, nucleic acids, etc.) through the same bright field and dark field counting methodology. The microparticles serve multiple functions: carrying capture antibodies, enabling digital counting, and providing a consistent detection platform across different analytes, thereby reducing overall system complexity while maintaining high sensitivity
Solution Approach 2:
The invention creates optical copies of microparticles through bright field and dark field imaging. Instead of directly manipulating physical molecules, the system captures optical images of microparticles at different illumination conditions, digitally counts these images, and infers molecular concentration from the count ratios. This copying approach simplifies the detection process by replacing complex molecular manipulation with straightforward image acquisition and analysis
3Measurement precision
If single-molecule level detection is performed, then accuracy for low-abundance molecules is improved, but the required optical detection sensitivity is extremely high
Solution Approach 1:
The invention introduces microparticles as intermediary carriers that bind to signal molecules. Instead of directly detecting individual molecules which requires extremely high optical sensitivity, the system detects microparticles that have captured molecules. The microparticles serve as amplifiers, converting single-molecule events into detectable optical signals through their larger size and enhanced optical properties in bright and dark field microscopy
Solution Approach 2:
The invention transitions from detecting molecules in a three-dimensional solution phase to detecting microparticles on a two-dimensional plane through bright field and dark field imaging. This dimensional change allows the use of conventional microscopy techniques with lower optical sensitivity requirements compared to direct single-molecule detection, while maintaining the ability to quantify low-abundance molecules through digital counting
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 stability, sensitivity, and accuracy of biomolecule detection while reducing costs, facilitating broader applications in scientific research, clinical diagnosis, and epidemic prevention.
Implementation Method 1
a common solid phase support (such as a glass slide, a multi-well plate and a flow channel) for random uniform distribution of microparticles
Implementation Method 2
direct imaging under bright and dark fields
Implementation Method 3
direct imaging under bright and dark fields
Data Source
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AI summary
The present invention provides a method and an apparatus for detecting molecules. The method for detecting a signal molecule comprises the following steps: (1) providing a solution comprising microparticles, wherein the microparticles comprise microparticles binding to the signal molecule to be detected; (2) applying the microparticles in the solution to the surface and/or the interior of a solid phase support; (3) counting the microparticles in a selected field of view under a bright field; (4) counting the microparticles binding to the signal molecule in a selected field of view under a dark field; and (5) determining the concentration of the signal molecule according to the counting results obtained in step (3) and step (4). On this basis, the present invention also provides a method and an apparatus for detecting a target molecule. The methods and apparatus provided by the present invention can realize a rapid, simple and convenient detection of molecules, especially biomolecules, which are of low cost, and facilitate promotion in multiple fields including scientific research, clinical diagnosis, and epidemic prevention.