Multi-Wavelength Optical pH Measurement for Cell Culture

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Solution Overview

Problem

Existing methods for measuring the pH of cell culture solutions, such as those containing fetal bovine serum (FBS), face errors due to visual checks and contamination risks with pH electrodes, and fail to accurately account for unknown FBS concentrations, leading to incorrect pH readings.

Innovation Solution

A pH measuring method using multiple light beams of specific wavelengths to illuminate the solution, measuring absorbances, and calculating pH based on these measurements, accounting for the absorbance characteristics of both phenol red and FBS, with expressions that include absorption coefficients and concentration calculations to correct for FBS influence.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Ease of manufacture

If visual check of phenol red color change is used to measure pH, then the measurement can be performed without specialized equipment, but measurement precision deteriorates due to errors and interference from serum yellow coloration

Engineering Contradiction:
Improvemeasurement simplicityVSAvoidpH measurement accuracy
Core Design Contradiction:
Ease of manufactureVSMeasurement precision

Solution Approach 1:

The patent replaces the visual/mechanical checking method with an optical measurement system using light emitting elements and light receiving elements. This substitution enables objective, quantitative pH measurement by detecting absorbance changes at specific wavelengths, eliminating both the simplicity of visual inspection and its associated errors and color interference issues.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

2Measurement precision

If pH electrodes are immersed in cell culture solution for measurement, then measurement precision can be improved with direct electrical measurement, but reliability deteriorates due to contamination risks from insufficient sterilization

Engineering Contradiction:
ImprovepH measurement accuracyVSAvoidsterility maintenance
Core Design Contradiction:
Measurement precisionVSReliability

Solution Approach 1:

The patent replaces the electrical measurement system (pH electrodes requiring sterilization) with an optical measurement system using light emitting and light receiving elements. This substitution maintains measurement precision while eliminating the sterilization requirements and contamination risks associated with immersing physical electrodes in the cell culture solution.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

3Device complexity

If conventional optical measurement at single wavelength is used, then device complexity can be reduced, but measurement precision deteriorates because FBS absorbance cannot be distinguished from phenol red absorbance

Engineering Contradiction:
Improvemeasurement system simplicityVSAvoidpH measurement accuracy
Core Design Contradiction:
Device complexityVSMeasurement precision

Solution Approach 1:

The patent segments the optical measurement into multiple discrete wavelength channels (411 nm for FBS, 430/560 nm for phenol red peaks, and reference wavelengths). By measuring absorbance at multiple separated wavelengths and processing the data separately, the system can distinguish and subtract FBS absorbance from phenol red absorbance, achieving accurate pH measurement while maintaining reasonable device complexity.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent extends the measurement from a single wavelength dimension to multiple wavelength dimensions. By adding the wavelength dimension and measuring absorbance spectra across multiple discrete wavelengths, the system creates an additional degree of freedom that enables separation of the overlapping absorbance signals from FBS and phenol red through spectral analysis.

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

4Measurement precision

If multiple light beams of different wavelengths are used to illuminate the solution, then measurement precision can be improved by accounting for FBS absorbance, but device complexity increases due to multiple light emitting elements and filtering requirements

Engineering Contradiction:
ImprovepH measurement accuracyVSAvoidoptical system complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent segments the optical system into distinct wavelength-specific components: light emitting elements for specific wavelengths (411 nm, 430 nm, 560 nm, and reference wavelengths), corresponding light receiving elements, and filters. This segmentation allows each component to be optimized for its specific wavelength range, achieving high measurement precision while keeping the overall device complexity manageable through modular design.

Inventive Principle:
Principle #1Segmentation

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 accurate pH measurement in cell culture solutions regardless of FBS concentration, reducing errors and contamination risks, and providing reliable pH readings within the suitable range for cell proliferation.

Implementation Method 1

measuring absorbances at the wavelengths of the received transmitted or reflected light beams of the first to fourth light beams respectively

Methodology Applied
Scientific EffectAbsorbance: Absorption (EM radiation)

Data Source

PatentEP2505992B1Method and apparatus for measuring the pH of a medium solution
Publication Date: 2018.09.19 NIHON KOHDEN CORP
  • EP2505992B1 patent drawingFigure 1
  • EP2505992B1 patent drawingFigure 2
  • EP2505992B1 patent drawingFigure 3

AI summary

A pH measuring method includes: illuminating a medium solution, which includes: a first material; and a second material, with a plurality of light beams, which includes: a first light beam, a wavelength of which corresponds to a first absorption peak; a second light beam, a wavelength of which corresponds to a second absorption peak or a second convergence point; a third light beam, a wavelength of which corresponds to a third absorption peak or a third convergence point; and a fourth light beam, a wavelength of the fourth light beam at which an absorbance of at least one of the first and second materials is converged irrespective of a pH; receiving transmitted or reflected light beams of the first to fourth light beams; measuring absorbances at the wavelengths of the received light beams respectively; and calculating a pH of the medium solution based on the absorbances.