Cell Migration Measurement Using Pillar Array Invasion Ratio
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Solution Overview
Problem
Conventional methods for measuring cell migration, such as those using micropillar chips, face challenges in distinguishing the degree of cell migration and invasion due to variations in cell adhesion and initial extracellular matrix positioning, leading to inaccurate measurement of cell migration distance.
Innovation Solution
A method involving the formation of cell spheroids on a pillar array within a well structure, where cells invade the micropillars, allowing for the calculation of an invasion ratio through fluorescence imaging, which accurately measures the extent of cell migration and invasion by differentiating between total cell area and spheroid area.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If cells are allowed to adhere to the contact surface of micropillars using conventional methods, then cell migration can be measured, but the initial position of cells in the extracellular matrix varies, causing different actual migration distances and reducing measurement accuracy
Solution Approach 1:
The invention segments the cell population into two distinct groups: cells that remain in the extracellular matrix (non-migrating) and cells that adhere to the micropillar contact surfaces (migrating). This segmentation allows for clear distinction and accurate measurement of cell migration by spatially separating migrating and non-migrating cells, directly resolving the issue of variable initial positions affecting measurement accuracy.
2Difficulty of detecting and measuring
If cell adhesion is used as the basis for measuring cell migration, then migration can be detected, but it becomes impossible to distinguish the degree of cell migration and invasion when the extent of adhered area varies
Solution Approach 1:
The invention employs fluorescent staining to visually differentiate between migrating cells (adhered to micropillars) and non-migrating cells (remaining in extracellular matrix). The fluorescent signal provides a clear visual and quantitative distinction, enabling accurate measurement of invasion degree without confusion from varying adhered areas, thus resolving the measurement precision issue.
3Productivity
If conventional micropillar chip methods are used to measure cell migration, then migration frequency can be measured, but variations in cell adhesion and initial extracellular matrix positioning lead to inaccurate measurement of cell migration distance
Solution Approach 1:
The invention replaces the conventional mechanical tracking method (tracking cell position changes over time) with a fluorescent staining-based detection method. This substitution allows for direct visualization and quantification of migrated cells through fluorescence intensity, eliminating the inaccuracies associated with mechanical position tracking and providing more precise migration distance measurement while maintaining high measurement efficiency.
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
This method enables precise measurement of cell migration and invasion by forming spheroids on a pillar array, allowing for the calculation of an invasion ratio, thereby overcoming the limitations of previous technologies in accurately assessing cell migration and invasion.
Implementation Method 1
staining and scanning the cell spheroids, the cells contained in the cell spheroids, and the cells that invaded the end contact surfaces of the micropillars
Data Source
AI summary
The present invention is directed to a method of measuring cell migration by measuring the invasion ratio of cells incubated on a pillar array inserted into a well structure, the method including steps of: preparing a pillar array having a plurality of micropillars and a well structure having a plurality of microwells into which the plurality of micropillars is insertable, respectively; forming cell spheroids by incubating cells in an extracellular matrix attached to the end contact surfaces of the micropillars; allowing the cells contained in the cell spheroids to invade the end contact surfaces; staining and scanning the cell spheroids, the cells contained in the cell spheroids, and the cells that invaded the end contact surfaces; and calculating the invasion ratio of cells by the following equation through a fluorescence image of the scanned cells:Invasion Ratio=Invasion cell areaTotal cell area=ATotal-AspheroidATotal[Equation]wherein Atotal represents the total cell area, and Aspheroid represents the spheroid area.