Spheroid Cell Death Analysis via Optical Tomography

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Solution Overview

Problem

Current methods fail to non-invasively and accurately assess cell death within spheroids, which are crucial for regenerative medicine, as conventional techniques cannot account for the three-dimensional structure and signal attenuation in spherical tissues, leading to inaccurate brightness comparisons and ineffective evaluation of cell death.

Innovation Solution

A method and apparatus utilizing optical imaging with reflected light to acquire cross-sectional images at varying distances, correcting brightness based on the three-dimensional shape and signal strength attenuation, allowing for non-invasive and quantitative analysis of cell death within spheroids.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If tissue staining is used to evaluate cell death in spheroid interior, then cell death can be detected, but the method is invasive and the evaluated spheroid cannot be used for transplantation

Engineering Contradiction:
Improvecell death detection accuracyVSAvoidinvasiveness of evaluation method
Core Design Contradiction:
Measurement precisionVSObject-affected harmful factors

Solution Approach 1:

The patent replaces the mechanical/chemical staining process with an optical measurement system. The light measurement device shines light through the spheroid and detects transmitted light intensity, substituting the invasive staining mechanism with a non-contact optical detection method that preserves spheroid viability for transplantation

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

Solution Approach 2:

The patent introduces light as an intermediary substance to detect cell death. Instead of using stains that physically interact with cells, light passes through the spheroid and its transmission characteristics reveal internal cell states, enabling non-invasive detection while maintaining spheroid integrity

Inventive Principle:
Principle #24Intermediary (Mediator)

2Object-affected harmful factors

If phase-contrast microscope observation is used to evaluate spheroid, then the method is non-invasive, but it cannot measure three-dimensional shape and evaluate cell death in spheroid interior

Engineering Contradiction:
Improvenon-invasive propertyVSAvoidthree-dimensional cell death evaluation capability
Core Design Contradiction:
Object-affected harmful factorsVSMeasurement precision

Solution Approach 1:

The patent transitions from two-dimensional phase-contrast microscopy to three-dimensional optical tomography. By measuring light transmission at multiple positions and reconstructing internal structures, the system achieves three-dimensional visualization of spheroid interior, enabling depth-resolved cell death evaluation while maintaining non-invasive operation

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

Solution Approach 2:

The patent creates a multi-functional measurement system that simultaneously performs three-dimensional shape measurement and internal cell death evaluation. The light measurement device not only captures the spheroid's external geometry but also detects internal cell states through transmitted light intensity variations, combining multiple evaluation capabilities in one non-invasive tool

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

3Device complexity

If conventional brightness comparison method is used in optical imaging, then the process is simple, but it produces inaccurate results due to signal attenuation in three-dimensional spherical tissue

Engineering Contradiction:
Improveimage analysis simplicityVSAvoidbrightness comparison accuracy
Core Design Contradiction:
Device complexityVSMeasurement precision

Solution Approach 1:

The patent replaces the simple but inaccurate conventional brightness comparison with a physics-based optical model. By substituting the direct comparison method with light transmission calculation that accounts for path length and tissue properties, the system achieves accurate cell death quantification while maintaining computational efficiency through automated algorithms

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

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 non-invasive assessment of cell death within spheroids, improving the quality of regenerative tissues by providing a direct evaluation of cell state, suitable for transplantation, and automating the analysis process.

Implementation Method 1

a light measurement device, irradiates the spheroid with light, and obtains a plurality of spheroid cross-sectional images at different distances from a light source position

Methodology Applied
Scientific EffectLight transmission: Light

Implementation Method 2

correcting brightness of the images in consideration of signal strength attenuation

Methodology Applied
Scientific EffectSignal attenuation: Absorption (EM radiation)

Data Source

PatentEP3473727B1Method for analyzing state of cells in spheroid
Publication Date: 2022.01.19 HITACHI HIGH TECH CORP
  • EP3473727B1 patent drawingFigure 1
  • EP3473727B1 patent drawingFigure 2
  • EP3473727B1 patent drawingFigure 3

AI summary

The present invention provides a method and an apparatus for analyzing a cell state, cell death in particular, in an interior of a spheroid non-invasively and quantitatively when the spheroid is cultured. More specifically, the present invention provides a method and an apparatus for analyzing a cell state by implementing optical imaging of a spheroid by using an optical instrument characterized by a high resolution and analyzing the internal structure of the spheroid.