Multi-Focal-Plane Cell Viability Determination Without Staining
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Solution Overview
Problem
Existing cell life-and-death determination techniques, such as those disclosed in JP2013-517460A, are insufficient for accurately distinguishing between alive and dead cells based on optical properties.
Innovation Solution
A method and device that utilize optical properties by capturing cell images in multiple focal planes, creating connected images, and analyzing feature amounts like lens effect, average refractive index, and diameter to determine cell viability, optionally using machine learning for improved accuracy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional cell life-and-death determination techniques are used, then the process is simple, but the measurement precision is insufficient to accurately distinguish between alive and dead cells
Solution Approach 1:
The imaging process is segmented into multiple focal planes (in-focus plane and out-of-focus planes) to capture comprehensive cell optical properties. This segmentation allows extraction of multiple feature amounts (lens effect, average refractive index, diameter) from different focal depths, improving measurement precision without requiring overly complex single-step imaging systems
Solution Approach 2:
The system transitions from single-plane imaging to multi-focal-plane imaging, adding the dimension of focal depth. By capturing images at different focal planes and creating connected images that integrate information across these planes, the system extracts more comprehensive optical features for more accurate cell viability determination
2Measurement precision
If staining reagents are used for cell determination, then the measurement precision improves, but the loss of substance increases and the ease of operation decreases
Solution Approach 1:
The system utilizes the cell's own optical properties (lens effect, refractive index, diameter) as natural markers for viability determination. By imaging and analyzing these inherent optical characteristics across multiple focal planes, the system eliminates the need for external staining reagents, achieving both high measurement precision and zero reagent consumption
Solution Approach 2:
The patent replaces chemical staining methods with optical imaging and analysis. Instead of using chemical reagents to mark viable cells, the system uses optical properties and machine learning algorithms to automatically distinguish alive from dead cells, eliminating reagent requirements while maintaining or improving measurement precision
3Measurement precision
If multiple focal plane images are captured and connected images are created, then the measurement precision improves, but the loss of time increases due to multiple imaging steps
Solution Approach 1:
The system performs preliminary automated processing of multi-focal-plane images by automatically creating connected images and extracting feature amounts (lens effect, average refractive index, diameter) without requiring manual intervention at each step. This preliminary automation reduces the time penalty of multi-plane imaging by streamlining the processing workflow
Solution Approach 2:
The system uses machine learning models that have been pre-trained on cell optical properties to rapidly analyze captured images and determine cell viability. The feedback mechanism allows the system to quickly process multiple focal planes by comparing extracted features against learned patterns, significantly reducing analysis time while maintaining high measurement precision
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 precise differentiation between alive and dead cells without staining reagents, facilitating efficient cell selection and concentration determination in cell suspensions.
Implementation Method 1
acquiring images of a cell captured in a plurality of focal planes including an in-focus plane of the cell
Implementation Method 2
extracting a feature amount from the connected image for analysis; the feature amounts include one or more feature amounts selected from a group consisting of a feature amount related to a lens effect of the cell
Implementation Method 3
a feature amount related to an average refractive index of the cell
Data Source
AI summary
Provided are a cell life-and-death determination method, a cell life-and-death determination device, and a cell life-and-death determination system. The cell life-and-death determination method includes: acquiring images of a cell captured in a plurality of focal planes including an in-focus plane of the cell in a direction opposite to a side on which the cell is irradiated with light; acquiring an image piece including a central portion and an outer peripheral portion of the cell from each of the images; connecting the image pieces in an order of an imaging direction of the focal plane to create a connected image for analysis; extracting a feature amount from the connected image for analysis; and determining whether the cell is alive or dead on the basis of the feature amount of the connected image for analysis and a predetermined range of the feature amount.