Reflection Quantitative Phase Microscopy for Stem Cell Flatness
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Solution Overview
Problem
Current methods for determining the differentiation level of induced Pluripotent Stem cells (iPS cells) are invasive, non-quantitative, and risk contamination, as they rely on staining or external reagents that can affect cell behavior, making them unsuitable for regenerative medicine applications.
Innovation Solution
A noninvasive method using the flatness of the cell surface as an indicator to determine differentiation level, where undifferentiated cells are flatter than differentiated cells, employing reflection quantitative phase microscopy to measure three-dimensional cell shape with high resolution, allowing for accurate differentiation level assessment without staining or external reagents.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If staining methods or external reagents are used to determine differentiation level, then measurement precision is improved, but object-affected harmful factors worsen due to cell sensitivity and contamination risks
Solution Approach 1:
The patent replaces chemical staining methods with physical optical measurement (reflection quantitative phase microscopy) to determine cell flatness. This substitution eliminates the need for external reagents that could contaminate cells or interfere with their differentiation state, while still providing quantitative measurement capability through optical phase difference detection
Solution Approach 2:
The patent introduces cell surface flatness as an intermediary parameter that correlates with differentiation level. By measuring flatness through non-invasive optical methods rather than directly measuring differentiation markers, the system avoids direct contact with cells using potentially harmful reagents while still obtaining accurate differentiation information
2Ease of operation
If phase contrast microscopy is used for visual observation, then ease of operation is improved, but measurement precision deteriorates due to lack of quantitativity
Solution Approach 1:
The patent replaces subjective visual observation with objective quantitative optical measurement. Reflection quantitative phase microscopy converts cell surface topography into quantifiable phase difference data, providing numerical measurements of flatness that correlate with differentiation level, thereby eliminating the subjectivity inherent in visual assessment
Solution Approach 2:
The patent changes the measurement parameter from qualitative visual appearance to quantitative flatness metrics (standard deviation of cell surface height, phase difference values). This parameter transformation enables precise numerical comparison of differentiation levels while maintaining the simplicity of optical observation
3Measurement precision
If fluorescent labeling or FACS methods are used, then measurement precision is improved, but device complexity and operation complexity worsen due to multiple reagents and equipment requirements
Solution Approach 1:
The patent replaces complex fluorescent labeling and FACS sorting equipment with a simpler reflection quantitative phase microscopy system. This substitution maintains measurement precision by using optical phase differences to detect cell flatness changes, while eliminating the need for fluorescent reagents, incubation steps, and complex flow cytometry instrumentation
Solution Approach 2:
The patent extracts only the essential measurement function (detecting cell flatness changes) from the complex staining and sorting process. By focusing solely on optical phase measurement of cell morphology, the system removes unnecessary reagents, equipment, and procedural steps while retaining the ability to distinguish differentiated from undifferentiated cells
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 effective and accurate determination of differentiation levels in iPS cells, facilitating quality control and improving their suitability for regenerative medicine by avoiding contamination risks and maintaining cell integrity.
Implementation Method 1
a method in which three-dimensional shapes of a plurality of cells are measured with use of a reflection quantitative phase microscope
Implementation Method 2
reflection quantitative phase microscope
Data Source
AI summary
The present invention provides a method for determining differentiation level of pluripotent stem cell, comprising a step of determining a flatness of cultured pluripotent stem cell, wherein the flatness is an indication.