Cell Cage Array for Single-Cell Isolation and Analysis
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Solution Overview
Problem
Current cytometry techniques face limitations in cell isolation, analysis, and manipulation, including overlapping fluorescent signals, inability to associate immunocytochemistry signals with individual cells, and dilution of cell lysate, which hinders precise analysis and monitoring of cellular changes.
Innovation Solution
A cell cage array apparatus that isolates cells using suction to draw them into cages with specific opening sizes, allowing only one cell per cage, and a conducting gel lid to trap cells, enabling individual cytometric analysis and ion injection without fluid volume change, facilitating direct quantitative analysis and multiple immunochemistry experiments.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If cells are analyzed by flowing them serially through a detector, then individual cell analysis is enabled, but fluorescent signals overlap and cannot be precisely distinguished
Solution Approach 1:
The invention divides the cell population into separate compartments by trapping individual cells in discrete cages within a gel matrix. Each cage acts as an isolated reaction chamber, preventing fluorescent signals from different cells from overlapping. This segmentation enables precise measurement of fluorescent signals from individual cells without the information loss that occurs in flow cytometry where signals from multiple cells overlap in time and space.
2Ease of operation
If cells are immobilized for analysis, then individual cell manipulation is enabled, but cell lysate becomes diluted and direct quantification is hindered
Solution Approach 1:
The gel cage structure creates localized compartments with distinct properties. Each cage provides a confined space that maintains high local concentration of cell lysate while still allowing the cell to be manipulated and analyzed. The small volume of each cage (on the order of picoliters) ensures that when a cell is lysed, the resulting lysate remains highly concentrated, enabling direct quantification without the dilution problems that would occur in larger analysis chambers.
3Measurement precision
If a gel layer is used to isolate cells, then individual cell trapping is achieved, but device complexity increases
Solution Approach 1:
The gel serves as an intermediary material that simplifies the overall device architecture. By using a gel matrix with embedded cages, the system achieves precise cell isolation without requiring complex mechanical trapping mechanisms, microfluidic channels, or multiple moving parts. The gel-cage combination acts as a passive, static structure that naturally confines cells through its physical properties, reducing device complexity while maintaining high measurement precision.
4Productivity
If multiple cells are present in analysis, then throughput is increased, but association of immunocytochemistry signals with individual cells is lost
Solution Approach 1:
The gel cage array simultaneously segments multiple cells into separate, identifiable compartments. Each cage acts as a discrete unit that can be individually addressed and analyzed. This segmentation enables high throughput analysis of many cells while preserving the association between each cell and its immunocytochemistry signals, because the spatial separation in the gel matrix allows each cell-cage pair to be tracked and analyzed independently, unlike bulk analysis methods where cell-signal associations are lost.
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 isolation and analysis of individual cells, allowing for serial immunochemistry experiments and direct quantification of cell lysate without dilution, overcoming limitations of existing cytometry methods.
Implementation Method 1
A pump may be used to draw the liquid medium into the enclosed reservoir
Implementation Method 2
Each cell cage may comprise a large opening adjacent to the first cell reservoir, a cell enclosure, and one or more small opening adjacent to the enclosed reservoir
Implementation Method 3
a gel layer forming a contiguous barrier to the large openings, isolating each cell in each of the cell enclosures
Implementation Method 4
One or more first electrode, in electric contact with the liquid medium in the cell reservoir... a current flowing between the one or more first and second electrodes inject ions into the cell cages
Implementation Method 5
the one or more first electrode is in electric contact with the liquid medium on one side of the enclosed reservoir, and the second electrode is in electric contact with the liquid medium on the opposite side of the enclosed reservoir, such that ion injection into cell cages is by diffusion
Data Source
AI summary
According to an aspect of some embodiments, an apparatus for isolation and cytometric analysis of cells from a liquid medium is provided. The apparatus comprises a cell reservoir, for receiving a liquid medium containing a suspension of cells. The apparatus further comprises a cell cage array between the cell reservoir and an enclosed reservoir, each cell cage comprising a large opening adjacent to the first cell reservoir, a cell cage, and one or more small opening adjacent to the enclosed reservoir. A connected liquid medium pump may move the suspended cells into cell cages by flowing the liquid medium from the cell reservoir to the enclosed reservoir. The apparatus further comprises a mechanical element which when actuated, pushes the cell cage array against a gel layer, forming a contiguous barrier to isolate each cell in a cell cage, and allowing cytometric analysis of isolated cells.