Cell Sphere State Determination via Holographic Phase Imaging
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for evaluating the quality of cell spheres in three-dimensional cultures are invasive, time-consuming, and require significant manpower, as they often involve destructive processes or two-dimensional evaluations that are not suitable for spheres of varying sizes in a three-dimensional space.
Innovation Solution
A non-destructive determination method that generates a phase difference image from a hologram of the cell sphere and uses a shape index value to assess the sphere's state, allowing for the evaluation of survival rate, density, homogeneity, and outer shape based on the correlation between the total phase difference amount and the shape index value.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional two-dimensional culture method is applied for evaluation, then evaluation can be performed, but the number of evaluation steps increases requiring much manpower and time
Solution Approach 1:
The patent replaces manual mechanical evaluation processes with automated optical imaging and image analysis. A camera captures images of cell spheres, and computer algorithms automatically analyze morphology, size, and other parameters, eliminating the need for manual counting and measurement while maintaining evaluation accuracy.
Solution Approach 2:
The patent creates visual copies (images) of cell spheres that can be analyzed without physically manipulating or destroying the original samples. Multiple images from different angles and focal planes are processed to extract quantitative data, enabling rapid assessment without time-consuming manual intervention.
2Measurement precision
If destructive treatment such as decomposing sphere into single cell or adding fluorescent coloring agent is performed, then evaluation can be achieved, but the evaluation process becomes complex and time-consuming
Solution Approach 1:
The patent enables cell spheres to be evaluated in their native state without requiring decomposition or staining. The imaging system captures optical properties and morphological features directly, allowing the spheres to 'serve themselves' as evaluation samples without additional processing steps.
Solution Approach 2:
The patent evaluates cell spheres by measuring optical parameters (light absorption, reflection, transmission) and morphological parameters (size, shape, surface texture) that naturally vary with cell state. These parameter changes provide diagnostic information without requiring chemical or physical modification of the samples.
3Loss of information
If conventional methods are used for spheres of various sizes in three-dimensional space, then evaluation is possible, but it is difficult to directly observe density and survival situation of cells inside the sphere
Solution Approach 1:
The patent transitions from two-dimensional surface observation to three-dimensional internal structure analysis by capturing images at multiple focal planes and processing them to reconstruct internal cell distribution, density, and spatial arrangement within the spheres.
Solution Approach 2:
The patent uses optical imaging as an intermediary to indirectly observe internal cell states. By analyzing light interaction patterns, morphological features, and density variations in captured images, the system infers internal cell survival and distribution without direct visual access to the interior.
4Productivity
If three-dimensional culture method is used for mass production of cells, then production efficiency increases, but non-destructive evaluation method for spheres has not been established
Solution Approach 1:
The patent replaces complex manual evaluation procedures with automated image capture and analysis systems. The camera and computer algorithms rapidly process multiple spheres simultaneously, providing quantitative assessment data without the time-consuming manual steps required by conventional methods.
Solution Approach 2:
The patent enables continuous evaluation of cell spheres throughout the culture process. The imaging system can assess spheres at various growth stages without interrupting culture conditions or requiring sample removal, maintaining continuous production flow while providing quality control data.
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 simple and accurate non-destructive evaluation of cell sphere quality, reducing labor and time while providing precise assessments of cell state and culture lot quality without the need for destructive treatments.
Implementation Method 1
imaging a sphere with an imaging unit to acquire a hologram of the sphere
Implementation Method 2
generating a phase difference image of the sphere from the hologram obtained by imaging the sphere
Data Source
Figure 1
Figure 2A~2D
Figure 3~4
AI summary
Provided is a determination method capable of non-destructively and simply determining a state of a sphere that is an aggregate of a plurality of cells. A phase difference image of a sphere that is an aggregate of a plurality of cells is generated from a hologram obtained by imaging the sphere, and a state of the sphere is determined on the basis of the phase difference image and a shape index value corresponding to a shape of the sphere.