Videomicroscopy Quantification Using Absorption Matrices

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Solution Overview

Problem

Current colorimetric analysis techniques in microscopy face limitations such as spectral overlapping, limited spectral resolution, and subjectivity in detecting and quantifying molecular species, leading to inaccurate results and high variability in sample analysis.

Innovation Solution

A method and system for refining the detection and quantification of molecular species using a videomicroscopy system that involves acquiring multiple images, determining optical density, and optimizing wavelength filter combinations to accurately calculate dye concentrations through the use of a relative absorption coefficient matrix and Cayley-Menger Determinants, thereby reducing errors and enhancing accuracy.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If colorimetric analysis techniques are used to detect and quantify molecular species, then the analysis can be performed rapidly and at low cost, but the results suffer from spectral overlapping, limited spectral resolution, and subjectivity leading to inaccurate quantification

Engineering Contradiction:
Improveanalysis speed and cost efficiencyVSAvoidquantification accuracy
Core Design Contradiction:
ProductivityVSMeasurement precision

Solution Approach 1:

The patent segments the spectral analysis by acquiring multiple images at different wavelength bands and processing them separately through optical density calculation and matrix inversion, thereby resolving spectral overlapping issues while maintaining rapid analysis capability

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent transitions from single-image colorimetric analysis to multi-image spectral analysis across multiple wavelength dimensions, using matrix mathematics to resolve the quantitative relationships between different molecular species and their absorption characteristics

Inventive Principle:
Principle #17Another dimension (Dimensionality change)

2Ease of operation

If standard colorimetric microscopy is used, then the system remains simple and easy to operate, but it cannot resolve spectral overlapping and provides limited spectral resolution

Engineering Contradiction:
Improvesystem simplicityVSAvoidspectral resolution
Core Design Contradiction:
Ease of operationVSMeasurement precision

Solution Approach 1:

The patent makes the microscopy system multi-functional by enabling it to perform both standard colorimetric analysis and enhanced spectral analysis using the same hardware platform, allowing users to choose the appropriate analysis depth based on their needs

Inventive Principle:
Principle #6Universality (Multi-functionality)

Solution Approach 2:

The patent changes the analysis parameters by introducing multiple wavelength filter combinations and using matrix inversion algorithms to extract quantitative information, thereby improving spectral resolution without requiring complex hardware modifications

Inventive Principle:
Principle #35Parameter changes

3Measurement precision

If multiple wavelength filters are used to improve quantification accuracy, then spectral overlapping is reduced, but the device complexity and computational requirements increase

Engineering Contradiction:
Improvedye concentration accuracyVSAvoidsystem complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent performs preliminary actions by pre-determining the relative absorption coefficient matrices for different molecular species and preparing the computational framework in advance, which simplifies the actual quantification process and reduces real-time computational complexity

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The patent creates a mathematical model (absorption coefficient matrix) that represents the physical system, allowing complex spectral relationships to be resolved through computational copying and manipulation of the matrix data rather than direct physical measurement

Inventive Principle:
Principle #26Copying

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 provides more accurate and reliable quantification of molecular species by minimizing errors due to noise fluctuations and spectral overlapping, resulting in improved analysis consistency and efficiency.

Implementation Method 1

determining an optical density of the sample in each pixel at each corresponding pixel location in the plurality of images

Methodology Applied
Scientific EffectOptical density measurement: Absorption Spectroscopy

Implementation Method 2

each molecular specie being indicated by a dye... determining an amount of each molecular specie, as indicated by a respective dye

Methodology Applied
Scientific EffectLight absorption by dyes: Absorption (EM radiation)

Data Source

PatentEP2697737B1Method for optimization of quantitative video-microscopy and associated system
Publication Date: 2019.07.31 TRIPATH IMAGING INC
  • EP2697737B1 patent drawingFigure 1
  • EP2697737B1 patent drawingFigure 2
  • EP2697737B1 patent drawingFigure 3

AI summary

Embodiments of the present invention are directed to a method for determining an amount of a plurality of molecular species in a sample, each molecular specie being indicated by a dye. According to one embodiment, the amount of a plurality of molecular species is determined by acquiring a plurality of images of the sample, determining an amount of each molecular specie, as indicated by a respective dye, for each pixel at each corresponding pixel location in the plurality of images, and refining the amount of a plurality of molecular species at one or more pixel locations in the plurality of images. Associated systems and computer program products are also provided.