Fluorescence Image Unmixing With Precomputed Separation Matrices
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Solution Overview
Problem
Conventional methods for acquiring separated images in multiple staining processes are hindered by increased calculation times and decreased throughput due to the type of excitation light and number of fluorescent dyes used.
Innovation Solution
A method involving the selection and generation of fluorescence separation images using image separation information based on reference fluorescence images in different optical states, utilizing a fluorescence filter unit with multiple reflection and transmission wavelength ranges, and excitation light beams of varying wavelengths to optimize image unmixing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional unmixing methods are used to separate fluorescence images, then separated images for each substance can be obtained, but calculation time increases and throughput decreases
Solution Approach 1:
The patent pre-calculates and stores unmixing coefficient matrices for multiple excitation light types and fluorescent dye combinations before actual imaging. During image acquisition, the system simply selects the pre-calculated matrix corresponding to the current excitation light and dye configuration, avoiding real-time complex calculations. This preliminary preparation significantly reduces calculation time during actual unmixing operations while maintaining separation accuracy.
Solution Approach 2:
The system pre-acquires reference fluorescence images under various excitation conditions and stores them in a database. When processing actual samples, the system retrieves appropriate reference images and uses pre-computed unmixing coefficients, eliminating the need to perform full unmixing calculations from scratch for each new sample, thereby improving throughput.
2Adaptability or versatility
If multiple excitation light types and fluorescent dyes are used for comprehensive observation, then more substances can be detected, but calculation time for unmixing increases
Solution Approach 1:
The patent segments the unmixing problem by pre-calculating separate unmixing coefficient matrices for different excitation light types and fluorescent dye combinations. Instead of performing one large, complex unmixing calculation for all possible combinations, the system divides the problem into multiple smaller, pre-solved cases. During actual operation, the system only needs to select and apply the appropriate pre-calculated matrix, dramatically reducing calculation time while maintaining the ability to handle diverse excitation and dye configurations.
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 reduces unmixing calculation amounts and improves throughput by leveraging pre-acquired image separation information, even with varying excitation lights and dye numbers.
Implementation Method 1
a fluorescence image is also acquired by irradiating the sample with excitation light
Implementation Method 2
through a fluorescence filter unit having a plurality of reflection wavelength ranges and a plurality of transmission wavelength ranges
Data Source
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AI summary
A fluorescent dye image acquisition system 1 includes an excitation light source 7 that irradiates a sample S with each of excitation light beams of a plurality of wavelength distributions; a camera 15 that acquires, for a plurality of types of fluorescence generated from the sample S by each of the excitation light beams of the plurality of wavelengths, target fluorescence images in a plurality of optical states having different wavelength characteristics through a camera-side filter set 9b; and an image processing device 5. The image processing device 5 stores a plurality of mixing matrices for obtaining fluorescent dye images of separated types of fluorescence, and generates a fluorescent dye image, for each of the plurality of types of fluorescence, based on the target fluorescence image that has been acquired in one optical state by the camera 15 and a selected mixing matrix.