Gaussian Residual Image Normalization for Particle Intensity
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Solution Overview
Problem
Current image processing methods in biotechnology, such as those using CCD detectors, face challenges in accurately normalizing images of particles, particularly in multiplexed applications, where fluorescent emission measurements can be affected by non-uniform light sources and spatial overlap of signals, leading to inconsistencies in characterizing cells or particles.
Innovation Solution
A method involving the acquisition of two-dimensional images of particles, where calibration particles are identified using a Gaussian or quadratic mathematical model to measure and normalize intensity, allowing for the correction of background signals and intensity variations across the image, thereby enhancing measurement accuracy and consistency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional image normalization methods are used, then processing speed is maintained, but measurement precision deteriorates due to non-uniform light sources and spatial overlap
Solution Approach 1:
The patent applies preliminary action by identifying and measuring calibration particles before normalizing the assay particles. The calibration particles are used to determine correction factors that are then applied to normalize the intensity measurements of assay particles, thereby correcting for non-uniform light sources and spatial overlap effects in advance
Solution Approach 2:
The calibration particles serve as an intermediary element between the light source and the assay particles. By measuring the intensity of calibration particles at different locations, the system derives correction factors that mediate the normalization process, allowing accurate measurement of assay particles despite non-uniform illumination and spatial overlap
2Measurement precision
If calibration particles are identified using mathematical models, then measurement precision improves, but processing time increases
Solution Approach 1:
The patent uses a simplified mathematical model (Gaussian or quadratic) to create a copy or representation of the calibration particle intensity profile. By fitting this simplified model to the measured intensity data, the system can quickly identify calibration particles and their locations without complex computation, thereby maintaining speed while improving precision
3Stability of the object's composition
If intensity normalization is performed, then measurement consistency improves, but device complexity increases
Solution Approach 1:
The patent changes the parameter of intensity measurement by applying correction factors derived from calibration particles. The normalization process adjusts the intensity values of assay particles based on the measured intensity and known properties of calibration particles, thereby stabilizing measurements across different locations and conditions without requiring complex hardware modifications
Data Source
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AI summary
An apparatus and method for image normalization using a Gaussian residual of fit selection criteria. The method may include acquiring a two-dimensional image of a plurality of particles, where the plurality of particles comprises a plurality of calibration particles, and identifying a calibration particle by correlating a portion of the image corresponding to the calibration particle to a mathematical model (e.g. Gaussian fit). The measured intensity of the calibration particle may then be used to normalize the intensity of the image.