In Situ Single Cell Lysis for Preserved Molecular Analysis
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Solution Overview
Problem
Current methods for analyzing single cells from solid tissues face challenges such as loss of cellular information due to tissue disaggregation, unpredictable molecular modification from fixation, and inability to construct standard curves, leading to inaccurate quantitative measurements of proteins, transcripts, and metabolites.
Innovation Solution
A method involving inside-out lysis using a detergent-containing buffer that spreads within the cell to preserve cellular components in an analytically defined state, allowing for the collection of lysate without fixation or disaggregation, which can be applied to analytical methods like mass spectrometry or microarrays.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If tissue disaggregation is used to obtain single cells, then cell suspension is achieved for analysis, but cellular information is lost and cell location data is unavailable
Solution Approach 1:
The patent extracts individual cells from solid tissue by targeting and lysing specific cells in place within the tissue architecture, rather than disaggregating the entire tissue. This allows extraction of cellular components from single cells while preserving spatial context and cellular information in the surrounding tissue environment.
2Stability of the object's composition
If fixation is applied to preserve tissue structure, then spatial distribution information is maintained, but molecular modification becomes unpredictable and quantitative measurement is compromised
Solution Approach 1:
The patent performs lysis of individual cells before any fixation or processing steps. By lysing cells in place within the tissue and collecting lysates directly, the method captures molecules in their native state without subsequent fixation-induced modifications, enabling accurate quantitative measurements while preserving spatial information through the in situ approach.
3Loss of information
If standard analytical methods are used on fixed tissue, then spatial distribution can be visualized, but construction of standard curves is impossible and quantitative accuracy is lost
Solution Approach 1:
The patent uses the tissue structure itself as an intermediary framework that preserves spatial relationships during the lysis process. By performing in situ lysis and collecting lysates while maintaining tissue architecture, the method enables both spatial visualization and quantitative analysis through standard curve construction, as the tissue serves as a spatial reference frame throughout the process.
4Productivity
If tissue disaggregation is performed to access single cells, then cell suspension is obtained, but cell yield is reduced and cell viability is compromised
Solution Approach 1:
The patent extracts cellular components directly from individual cells within the tissue by targeted lysis, rather than attempting to dissociate and suspend cells. This approach achieves complete cellular lysis and component extraction without the need for tissue disaggregation, thereby maximizing cell yield while preserving the viability and functionality of remaining cells in the tissue.
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 accurate, multiplexed measurement of native proteins, transcripts, and metabolites in single cells with preserved cellular information, overcoming limitations of existing methods by maintaining the natural state of cellular molecules and allowing for concurrent standard curve determination.
Implementation Method 1
contacting a detergent-containing lysis buffer with the intracellular space of the identified cell; allowing the lysis buffer to spread within the intracellular space
Data Source
AI summary
The invention provides a method for lysing a single cell embedded in a tissue from the inside of the cell, and collecting the intracellular lysate for use in analytical methods. This method preserves the state of molecules of the cell, and therefore allows for transformation of a single target cell in live tissue into a format that can be evaluated using analytical methods.