Cell Spheroid Protrusion Evaluation via Fluorescence Microscopy
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Solution Overview
Problem
Conventional cell invasion assays using plane-cultured cells cannot evaluate the migration/invasion ability of cells in a three-dimensional structure, which is crucial for mimicking the in vivo microenvironment of cancer tissues and assessing the effectiveness of anticancer agents.
Innovation Solution
A method involving imaging and analyzing cell spheroids labeled with fluorescent substances using fluorescence microscopy to determine protrusion-forming ability, allowing for the evaluation of cell spheroids in a three-dimensional structure, including steps for imaging, analyzing, and evaluating protrusions based on various parameters such as length, width, and branching.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If plane-cultured cells are used for cell invasion assay, then the assay can be performed using conventional methods, but the migration/invasion ability cannot be evaluated in a three-dimensional structure
Solution Approach 1:
The patent transitions from two-dimensional plane-cultured cells to three-dimensional cell spheroids, enabling evaluation of migration/invasion ability in a three-dimensional structure that better mimics in vivo conditions while maintaining assay feasibility
2Measurement precision
If three-dimensional cell spheroids are used to mimic in vivo microenvironment, then the evaluation accuracy improves, but the imaging and analysis complexity increases
Solution Approach 1:
The patent uses fluorescent substances that emit different colors/wavelengths to label cell spheroids and protrusions, enabling differentiation and quantitative analysis of structural features through fluorescence microscopy without requiring complex imaging systems
Solution Approach 2:
The patent replaces complex mechanical measurement systems with fluorescence-based optical imaging and image analysis, simplifying the measurement process while maintaining high evaluation accuracy for protrusion-forming ability
3Ease of operation
If conventional transmission light microscopy is used to image cell spheroids, then the imaging process is simple, but the resolution is insufficient to identify individual cells in three-dimensional structure
Solution Approach 1:
The patent employs fluorescent labeling that emits specific wavelengths of light, allowing high-resolution imaging of individual cells within three-dimensional spheroids through fluorescence microscopy while maintaining operational simplicity
Solution Approach 2:
The patent uses fluorescent substances as intermediaries to enhance the visibility and resolution of cell structures, enabling clear identification of individual cells and protrusions without complicating the imaging process
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 the quantitative evaluation of protrusion-forming ability of cell spheroids in a three-dimensional structure, providing insights into cancer cell invasion and sensitivity to anticancer agents, and simulating the in vivo microenvironment for more accurate assessments.
Implementation Method 1
imaging and analyzing cell spheroids labeled with fluorescent substances using fluorescence microscopy
Data Source
AI summary
The present invention provides a method for evaluating a protrusion-forming ability of cell spheroids, comprising step (a) of imaging a cell spheroid labeled with a fluorescent substance using a fluorescence microscope at a resolution capable of identifying individual cells, and acquiring a plane tomographic image of a fluorescence emitted from the fluorescent substance, step (b) of analyzing the plane tomographic image acquired in step (a) to determine a protrusion part of the cell spheroid, and step (c) of evaluating a protrusion-forming ability of the cell spheroid based on the protrusion part determined in step (b).