Fluorescence Imaging of Moving Particles via Iterative ROI Extraction
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Solution Overview
Problem
Current fluorescence imaging techniques are inadequate for observing moving particles, such as spermatozoa, due to their unpredictable trajectories and the need for prolonged acquisition times or image cumulation, which is unsuitable for particles moving along different paths.
Innovation Solution
A method involving illuminating a sample in a spectral detection band different from the emission band, forming detection and emission images, and iteratively extracting regions of interest to create an integrated emission image, which enhances the signal-to-noise ratio by interpolating particle positions and averaging fluorescence signals from multiple images.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If the acquisition time is increased to improve the signal-to-noise ratio of fluorescence images, then the signal-to-noise ratio is improved, but the particle moves along an unpredictable trajectory making the image useless for characterization
Solution Approach 1:
The patent segments the fluorescence image acquisition into multiple short-exposure images taken at different time points. Instead of taking one long exposure that would cause particle movement blur, the system takes multiple short exposures and processes them separately to reconstruct the particle's fluorescence signal while tracking its movement trajectory.
Solution Approach 2:
The patent performs preliminary detection of particle position and trajectory using the short-exposure images before final fluorescence image reconstruction. By predicting the particle's trajectory in advance based on detected positions, the system can correctly assign fluorescence signals to the appropriate particle locations in the reconstructed image, even though the particle moves during acquisition.
2Measurement precision
If multiple fluorescence images are cumulated to improve the signal-to-noise ratio, then the signal-to-noise ratio is improved, but each particle follows a different trajectory making cumulation ineffective
Solution Approach 1:
The patent implements a feedback mechanism where particle positions are detected in each short-exposure image, the trajectory is updated based on these detections, and this trajectory information is fed back into the fluorescence image reconstruction process. This feedback loop ensures that fluorescence signals are correctly assigned to the moving particle's position at each time point, enabling effective cumulation despite particle movement.
Solution Approach 2:
The patent changes the temporal parameter of image acquisition by using multiple short-exposure images instead of one long exposure. This parameter change allows the system to capture particle position information at different time points while maintaining the ability to reconstruct the fluorescence signal through trajectory-based assignment and cumulation.
3Speed
If short exposure time is used to capture moving particles, then the particle position is accurately captured, but the fluorescence signal intensity is too low for characterization
Solution Approach 1:
The patent merges multiple low-intensity fluorescence signals from short-exposure images into a single high-intensity reconstructed image. By detecting particle positions in each short exposure, tracking the trajectory, and assigning fluorescence signals to the correct positions along the trajectory, the system combines the weak signals from multiple exposures to create a usable fluorescence image that accurately represents the moving particle.
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 allows for effective observation of fluorescence or luminescence from moving particles by improving the signal-to-noise ratio through iterative image processing and integration, enabling accurate tracking and characterization of particles like spermatozoa.
Implementation Method 1
The particle emits a fluorescence light in the spectral emission band when it is illuminated in a spectral excitation band
Data Source
AI summary
Method for observing an emission of fluorescence light or luminescence light from a moving particle in a sample, in a spectral emission band, the method comprising:a) forming a detection image of the sample in a spectral detection band, the spectral detection band being different from the spectral emission band;b) forming an emission image of the sample in the spectral emission band;the method being such that:steps a) and b) are reiterated;the method comprising:c) on the basis of each detection image resulting from step a) of each iteration of steps a) and b), detecting the particle and determining a region of interest around the particle;d) on the basis of the region of interest resulting from each step c), extracting a region of interest from each emission image of the sample.