Single-Cell Capture Substrates for Multiplexed Secretion Profiling
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Solution Overview
Problem
Existing methods for bulk analysis of immune response are inadequate for evaluating crucial cellular interactions at the single-cell level, failing to measure protein secretion in a highly multiplexed manner or analyze cell-cell interactions, which hinders the development of effective therapeutic treatments.
Innovation Solution
Systems and devices for positioning, sealing, and isolating single cells, along with methods for manufacturing capture substrates, enable highly multiplexed analysis of single-cell secretion profiles by using compressible substrates with micro-chambers and capture antibodies, and instruments for imaging and analyzing expressed substances.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If bulk analysis methods are used to evaluate immune response, then the analysis process is simple, but the ability to evaluate cellular interactions at the single-cell level is lost
Solution Approach 1:
The system divides the sample into individual single cells using microfluidic droplet generation, where each droplet contains at most one cell. This segmentation enables single-cell level analysis while managing complexity through automated droplet formation and processing
Solution Approach 2:
The system implements nested structures where micro-chambers are embedded within a larger microfluidic device, and capture substrates are positioned within individual chambers. This nesting allows single-cell isolation and analysis while maintaining an integrated system architecture
2Adaptability or versatility
If single-cell analysis technologies are developed to understand immune response, then the understanding of cellular interactions improves, but the ability to directly measure protein secretion in a highly multiplexed manner remains limited
Solution Approach 1:
The capture substrate is designed with multiple capture areas, each functionalized with different capture antibodies to detect multiple different proteins simultaneously. This multi-functional design enables highly multiplexed protein secretion analysis from single cells while maintaining measurement precision through specific antibody-protein binding
Solution Approach 2:
The system uses capture antibodies as intermediaries between the secreted proteins and the detection system. These antibodies specifically bind to target proteins, enabling precise measurement of protein secretion while allowing multiplexed detection through different antibody specificities
3Reliability
If single-cell analysis is performed to evaluate cell-cell interactions, then the therapeutic treatment development improves, but the current technologies fail to analyze interactions at the single cell level
Solution Approach 1:
The system performs preliminary actions by isolating and positioning single cells in micro-chambers before analysis, and by pre-functionalizing capture substrates with specific antibodies. This preliminary preparation enables reliable cell-cell interaction analysis while managing complexity through standardized protocols and pre-configured components
Data Source
AI summary
Embodiments of the current disclosure are directed to systems, methods and apparatus for evaluating single cell secretion profiles. In some embodiments, the apparatus may be configured to analyze substances expressed by a biological cell and may include a first compressible substrate, and a second substrate configured for removable sealing attachment with the first substrate. In some embodiments, upon attachment of the second substrate with the first substrate, an assembly is formed such that the open side of the plurality of chambers are covered by the second substrate, and a portion of each of the plurality of capture areas are exposed in each of the chambers.