Imaging Flow Cytometry With Spatial Coding for Parallel Biomarker Assays
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Solution Overview
Problem
Existing detection methods for biological particles, such as flow cytometry and fluorescence microscopy, are limited in throughput and spatial resolution, unable to efficiently detect 10^2 to 10^5 different objects simultaneously, and spatially coded reagents smaller than 100 nm cannot be effectively resolved by imaging flow cytometry.
Innovation Solution
A method combining imaging flow cytometry with spatially coded reagents, using fluorescent microspheres of different colors and spatial positions to form coding sequences, enabling high-throughput parallel detection of biological particles, nucleic acids, proteins, and single cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional flow cytometry is used for high-throughput detection, then detection speed can reach 10^3 particles per second, but the number of parallel assays is limited to 50 different analytes due to spectral width constraints
Solution Approach 1:
The patent transitions from spectral coding (1D wavelength dimension) to spatial coding (2D position dimension) of fluorescent microspheres. By arranging microspheres with different fluorescence colors in specific spatial positions, the system achieves combinatorial coding capacity of 10^2 to 10^5 different analytes while maintaining high detection throughput, effectively resolving the contradiction between throughput and parallel assay capacity.
2Adaptability or versatility
If spatially coded reagents with smaller size are used to increase coding capacity, then more analytes can be detected in parallel, but detection accuracy decreases when size is below 100 nm
Solution Approach 1:
The patent optimizes the size parameter of fluorescent microspheres to be within 100-1000 nm, which is large enough to be effectively resolved by imaging flow cytometry (maintaining detection accuracy) while still enabling high coding capacity through spatial arrangement. This parameter optimization resolves the contradiction between coding capacity and detection accuracy.
3Adaptability or versatility
If more fluorescent labels with different colors are used to increase parallel detection capacity, then more analytes can be detected, but spectral overlap increases making differentiation difficult
Solution Approach 1:
The patent moves from relying solely on spectral differentiation (1D wavelength) to spatial-spectral combined coding (2D position + wavelength). By encoding information in both the spatial position and fluorescence color of microspheres, the system achieves high parallel detection capacity while maintaining accurate differentiation through the added spatial dimension, effectively resolving the spectral overlap problem.
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
Achieves high-throughput parallel detection of 10^2 to 10^5 biological particles per second, with improved accuracy and efficiency, suitable for liquid biopsy applications.
Implementation Method 1
selecting fluorescent labels having small fluorescence spectral overlap, labeling the surface of microspheres with the fluorescent labels
Data Source
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AI summary
A high-throughput parallel testing method based on imaging flow cytometry and spatially coded reagents combines the high-throughput characteristic of an imaging flow cytometer and the high coding capacity characteristic of spatially coded reagents, thereby achieving high-throughput parallel testing of nucleic acid sequencing, protein testing and single cell analysis, and being capable of being used for liquid biopsy or other biomedical use requirements for performing high-throughput synchronous testing on different biomarkers.