Live-bead Classification via Autofluorescence Correction
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Solution Overview
Problem
Current DNA sequencing technologies face challenges in distinguishing between live-beads (DNA-bound) and blank-beads due to weak fluorescent signals and the inability to accurately assess the enrichment ratio, requiring precise optical equipment and extensive image evaluation.
Innovation Solution
A computer-implemented method and system that corrects autofluorescence and crosstalk effects in digital images by determining intensity values, computing standard deviations, normalizing data, and applying classification thresholds to differentiate live-beads from blank-beads using a threshold selection method, specifically addressing issues of inhomogeneous image brightness and uneven illumination.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If fluorescent signals are used to detect DNA-bound beads, then object classification can be performed, but the signals are weak and require precise optical equipment
Solution Approach 1:
The patent applies preliminary action by performing background subtraction and signal normalization before final object classification. The system pre-processes the fluorescent images by removing background noise and normalizing signal intensities across different channels, which enhances the weak fluorescent signals from DNA-bound beads without requiring more complex optical equipment.
Solution Approach 2:
The patent uses computational image processing as an intermediary between the fluorescent detection and object classification. By introducing algorithms that perform background subtraction, signal normalization, and threshold-based classification, the system bridges the gap between weak fluorescent signals and reliable object identification without increasing optical complexity.
2Measurement precision
If extensive image evaluation is performed to obtain precise sequencing results, then classification accuracy improves, but processing time and computational resources increase
Solution Approach 1:
The patent applies partial action by performing image evaluation only on regions containing beads of interest, rather than processing the entire image uniformly. The system identifies candidate bead regions and applies detailed image evaluation algorithms selectively to these regions, reducing overall processing time while maintaining classification accuracy for relevant objects.
Solution Approach 2:
The patent segments the image processing task into distinct stages: background subtraction, signal normalization, threshold-based preliminary classification, and detailed evaluation only for candidate regions. This segmentation allows the system to perform extensive evaluation where needed while skipping unnecessary processing in other areas, balancing accuracy and processing time.
3Reliability
If white-light illumination is used to determine bead positions, then position determination is robust, but live-beads cannot be distinguished from blank-beads
Solution Approach 1:
The patent merges white-light illumination for position determination with fluorescent illumination for live-bead identification. The system first uses white-light images to robustly determine bead positions, then overlays fluorescent channel information at these positions to distinguish live-beads from blank-beads, combining the advantages of both illumination methods.
Solution Approach 2:
The patent transitions from a single white-light dimension to multiple fluorescent wavelength dimensions. By analyzing the same bead positions across multiple fluorescent channels with different emission spectra, the system extracts additional information that enables live-bead identification while maintaining the position determination accuracy provided by white-light illumination.
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
Enables reliable and robust identification of live-beads, improving the assessment of enrichment ratios and enhancing the accuracy of DNA sequencing results by overcoming previous limitations in signal detection and image analysis.
Implementation Method 1
The digital images are determined by an optical imaging system during emission of electromagnetic radiation by the fluorescent compounds
Data Source
AI summary
System and method for distinguishing at least one first object from at least one second object in a plurality of digital images is provided. The at least one first object having received at least one molecule comprising genetic information, the at least one second object not having received a molecule comprising genetic information. The at least one molecule is configured to receive one of a plurality of fluorescent compounds in each of a plurality of cycles. The digital images being determined by an optical imaging system during emission of electromagnetic radiation by the fluorescent compounds, wherein the plurality of digital images comprises a plurality of series of images, each image of a series referring to the emission spectrum of a respective fluorescent compound and wherein the series of images is repeatedly taken for each of the plurality of cycles.