Single Molecule Identification via Intensity Time Sequencing
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Solution Overview
Problem
Current single molecule sequencing technologies face challenges in accurately identifying and counting single molecules due to variations in fluorophore emission times, light intensities, and background noise, leading to labor-intensive and inefficient methods.
Innovation Solution
A method and device that process a time sequence of intensity data by forming a line chart, dividing it into grids, applying line erosion, and using run-length coding to identify and count single molecules based on predetermined thresholds, resulting in a simplified image for quick and accurate identification and counting.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If HMM and machine learning methods are used for single molecule identification, then identification capability is improved, but training time and operational efficiency deteriorate
Solution Approach 1:
The patent segments the continuous fluorescence intensity signal into discrete intensity levels through thresholding, dividing the signal space into distinguishable regions. This segmentation enables direct pattern recognition without requiring extensive machine learning training, thus improving operational efficiency while maintaining identification accuracy.
Solution Approach 2:
The patent creates simplified representations (copies) of the fluorescence signals by converting them into intensity level sequences and morphological features. These copied representations capture the essential characteristics needed for identification while eliminating the need for complex training processes associated with raw signal analysis.
2Adaptability or versatility
If manual identification methods are used, then flexibility and adaptability are improved, but labor cost and time consumption deteriorate
Solution Approach 1:
The patent implements self-service through automated image processing algorithms that perform morphology operations, intensity level determination, and molecule identification without human intervention. The system processes fluorescence images through a standardized workflow that maintains adaptability to different molecular types while dramatically improving productivity through automation.
Solution Approach 2:
The patent changes parameters by converting continuous fluorescence intensity values into discrete intensity levels and by transforming spatial image data into morphological features. These parameter transformations enable automated processing while preserving the information needed for accurate identification, thus improving productivity without sacrificing adaptability.
3Measurement precision
If fluorescence intensity detection is performed, then single molecule identification capability is improved, but identification accuracy deteriorates due to varying emission times and light intensities
Solution Approach 1:
The patent applies preliminary action by performing morphology operations and background subtraction before intensity measurement. These preprocessing steps normalize the fluorescence signals and remove artifacts that cause variability, ensuring that subsequent intensity-based identification is performed on standardized data, thus improving both accuracy and reliability.
Solution Approach 2:
The patent transitions from analyzing fluorescence in the time dimension to analyzing intensity levels in a discrete spatial dimension. By converting continuous temporal fluorescence signals into discrete intensity level sequences and applying morphology operations, the method creates a new dimensional representation that is less sensitive to variations in emission timing and light intensity, thereby improving identification consistency.
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 enables rapid and precise identification and counting of single molecules, reducing human error and increasing operational efficiency in gene sequencing processes.
Implementation Method 1
a base group is identified by fluorescence which is a light intensity emitted from an excited state to a ground state at a specific power laser irradiation
Data Source
AI summary
A method and a device for identifying a single molecule, a method and a device for counting a single molecule and a system for processing a single molecule are provided. The method for identifying the single molecule includes inputting time sequence of an intensity of an spot; forming a line chart of time and intensity of the spot according to the time sequence, wherein the line chart consists of a plurality of line segments; dividing the line chart into a plurality of grids in an array, counting the number of the line segments and/or ends of the line segments in each grid; subjecting the divided line chart to a line erosion according to numbers corresponding to the grids to transform the divided line chart to a simplified image; subjecting the simplified image to run-length coding to indicate connected domains; calculating an area of each connected domain and determining that the connected domain corresponds to a single molecule if the area of the connected domain is greater than a first predetermined threshold. According to the method for identifying the single molecule, the line chart of the time sequence of the intensity of the spot is processed to transform into the image, which results in a quick identification for the single molecule and a high accuracy of the identification.