Non-Invasive Cell Count Estimation via Metabolite Spectroscopy

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Solution Overview

Problem

Current methods for estimating the number of cells in culture, such as bioassays and spectroscopy, are invasive or limited in accuracy, especially when cellular metabolite concentrations are low, making continuous and non-invasive monitoring challenging.

Innovation Solution

A method and device that estimate the number of cells by switching between measuring cell-consumed and cell-produced substances using spectroscopy or electrochemical systems, applying thresholds to ensure accurate concentration estimation and applying relationship information to estimate cell numbers, allowing continuous and non-invasive monitoring.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If bioassay is used to obtain accurate relationship information between cellular metabolite concentration and cell number, then measurement precision is improved, but the method becomes invasive and cannot perform continuous monitoring

Engineering Contradiction:
Improveaccuracy of relationship informationVSAvoidcontinuous non-invasive monitoring capability
Core Design Contradiction:
Measurement precisionVSEase of operation

Solution Approach 1:

The patent segments the metabolite monitoring into multiple wavelength measurements. By measuring absorbance at multiple wavelengths (including isosbestic points), the system can differentiate between various metabolites and maintain accurate cell number estimation without invasive sampling throughout the culture period.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent replaces the mechanical/invasive bioassay sampling method with optical spectroscopy. The spectrometer measures metabolite concentrations non-invasively by detecting light absorbance, eliminating the need for physical sampling while enabling continuous monitoring.

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

2Ease of operation

If spectroscopy is used to enable continuous non-invasive monitoring, then ease of operation is improved, but measurement precision deteriorates when cellular metabolite concentration is low

Engineering Contradiction:
Improvecontinuous non-invasive monitoring capabilityVSAvoidaccuracy of cellular metabolite concentration estimation
Core Design Contradiction:
Ease of operationVSMeasurement precision

Solution Approach 1:

The patent transitions from single-wavelength absorbance measurement to multi-wavelength spectral analysis. By measuring absorbance across multiple wavelengths and applying multivariate analysis, the system can accurately distinguish metabolite concentrations even at low levels, overcoming the precision limitations of single-wavelength methods.

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

Solution Approach 2:

The patent uses a composite analytical approach combining measurements at multiple wavelengths with multivariate analysis algorithms. This composite method processes spectral data from various wavelengths simultaneously, improving the accuracy of metabolite concentration estimation when concentrations are low.

Inventive Principle:
Principle #40Composite materials

3Device complexity

If a single cellular metabolite is monitored throughout the culture period, then device complexity is reduced, but reliability deteriorates due to concentration changes affecting accuracy

Engineering Contradiction:
Improvesimplicity of monitoring systemVSAvoidconsistency of cell number estimation accuracy
Core Design Contradiction:
Device complexityVSReliability

Solution Approach 1:

The patent employs multiple metabolites (both consumed and produced substances) that serve different functions throughout the culture period. By monitoring multiple metabolites simultaneously, the system maintains reliable cell number estimation across all culture stages, with each metabolite providing accurate information during specific phases.

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

Solution Approach 2:

The patent implements a feedback mechanism where the system continuously monitors metabolite concentrations and uses this information to estimate cell numbers. The relationship information obtained from bioassays serves as a reference feedback to validate and calibrate the spectroscopic measurements, ensuring consistent accuracy throughout the culture period.

Inventive Principle:
Principle #23Feedback

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 and continuous non-invasive estimation of cell numbers throughout the culture period by leveraging the concentration of cell-consumed and cell-produced substances, improving upon the limitations of existing techniques.

Implementation Method 1

a culture medium is irradiated with light having a wavelength that does not adversely affect cells, and the absorbance and/or the degree of scattering thereof are measured

Methodology Applied
Scientific EffectAbsorbance: Absorption (EM radiation)

Implementation Method 2

adding a reagent that reacts only with a specific cellular metabolite to the collected media; and detecting a luminescence signal produced by reacting the reagent with the cellular metabolite in the media

Methodology Applied
Scientific EffectLuminescence: Luminescence

Data Source

PatentEP3859008B1Method for estimating cell count, and device for estimating cell count
Publication Date: 2022.11.16 PHC HLDG CORP
  • EP3859008B1 patent drawingFigure 1
  • EP3859008B1 patent drawingFigure 2
  • EP3859008B1 patent drawingFigure 3

AI summary

[Object] To provide a method and device for estimating the number of cells using a non-invasive process. [Solution] To estimate concentration of a cellular metabolite contained in a culture medium where certain cells are cultured, using a simple method, etc., spectroscopy. The number of the certain cells is estimatable by applying previously obtained information regarding relationship between a cellular metabolite concentration and the number of the certain cells. In spectroscopy, the higher the cellular metabolite concentration, the more accurately the actual concentration of the cell consumed-substance can be estimated. Accordingly, the concentration of a cell-consumed substance, decreasing as cells are cultured, are estimated in the early to middle stages of culture, and the concentrations of a cell-produced substance, increasing as cells are cultured, are estimated in the middle to late stage of the culture. This enables estimation of the number of cells in the entire range from beginning to end of cell culture.