Digital Holography for Non-Destructive 3D Cell Aggregate State Determination
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Solution Overview
Problem
Existing methods for evaluating the state of cell aggregates in three-dimensional cultures are destructive, time-consuming, and require significant manpower, especially when dealing with varying sizes, making it difficult to assess cell density and survival within spheres.
Innovation Solution
A non-destructive method using digital holographic microscopy to generate phase difference images of cell aggregates, deriving phase difference amount density, and analyzing its time transition to determine the state and differentiation of cells, including processes to differentiate stem cells into germ layers and specific cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional two-dimensional culture evaluation methods are applied to three-dimensional cell aggregates, then cell state can be evaluated, but the evaluation process becomes destructive and time-consuming
Solution Approach 1:
The patent replaces mechanical/destructive evaluation methods with optical measurement using digital holographic microscopy. The system captures holograms of cell aggregates and uses phase difference imaging to non-destructively assess cell density and state, eliminating the need for physical disruption or fluorescent labeling while maintaining evaluation accuracy
Solution Approach 2:
The patent creates optical copies (holograms) of the cell aggregates that contain information about cell density and state. By analyzing the phase difference in these optical copies, the system can evaluate the original cell aggregates without direct contact or destruction, enabling repeated measurements over time
2Loss of information
If conventional evaluation methods with fluorescent coloring agents are used, then cell information can be obtained, but the cells are destroyed and manual intervention is required
Solution Approach 1:
The patent substitutes chemical labeling (fluorescent coloring agents) with physical optical measurement. The digital holographic microscopy system extracts cell information through phase difference analysis of light passing through the aggregates, eliminating the need for complex staining protocols and manual image processing while preserving cell viability
Solution Approach 2:
The cell aggregates themselves provide the measurement signal through their inherent optical properties (refractive index differences). The system requires no external labels or markers - the cells' own physical characteristics are sufficient for evaluation, simplifying the process and avoiding destruction
3Quantity of substance
If the number of cells to be cultured increases in three-dimensional aggregates, then production capacity increases, but the number of evaluation steps increases requiring more manpower
Solution Approach 1:
The patent creates a universal evaluation system that handles cell aggregates of various sizes and densities through a single automated optical measurement process. The digital holographic microscopy system can evaluate multiple aggregates simultaneously or in sequence without requiring different methods for different cell numbers, eliminating the need for multiple evaluation steps and manual intervention
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 easy and accurate determination of cell aggregate states and differentiation ratios without destroying the cells, reducing time and manpower, and providing insights into cell health and behavior over time.
Implementation Method 1
generating a phase difference image of an aggregate of a plurality of cells from a hologram obtained by imaging the aggregate
Implementation Method 2
generating a phase difference image of an aggregate of a plurality of cells from a hologram obtained by imaging the aggregate
Data Source
Figure 1
Figure 2A~2D
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AI summary
A determination method of non-destructively and easily determining a state of an aggregate of a plurality of cells formed by three-dimensional culture is provided. A determination method according to the disclosed technology includes generating a phase difference image of an aggregate of a plurality of cells from a hologram obtained by imaging the aggregate, deriving a phase difference amount density by dividing a total phase difference amount that is a value obtained by integrating a phase difference amount of each of a plurality of pixels constituting the phase difference image by a volume of the aggregate, and determining a state of the aggregate on the basis of a time transition of the phase difference amount density.