Deep Eutectic Solvent Fixation for Single-Cell Dissociation
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Solution Overview
Problem
Current methods for dissociating biological samples into single cells fail to preserve the original morphology and biomolecule composition, leading to inaccurate representation of the transcriptome and proteome, which can result in incorrect therapeutic interventions for cancer and other diseases.
Innovation Solution
The use of Deep Eutectic Solvents (DES) for fixing tissues, followed by sonication to dissociate cells, which effectively stabilizes biomolecules and maintains cell morphology, allowing for detailed analytical procedures like single-cell RNA sequencing and flow cytometry.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If enzymatic treatment or mechanical action is used to dissociate samples into single cells, then cell separation is achieved, but cell morphology and biomolecule composition are not preserved
Solution Approach 1:
The patent applies preliminary fixation of the biological sample with a fixative (such as formaldehyde, methanol, or acetone) before dissociation into single cells. This preliminary action preserves the cell morphology and biomolecule composition in their native state, ensuring that subsequent dissociation methods (enzymatic or mechanical) do not alter the original cellular structures or molecular integrity, thus resolving the contradiction between achieving cell separation and preserving biomolecule accuracy
Solution Approach 2:
The patent changes the physical and chemical parameters of the dissociation process by controlling enzyme concentration, incubation time, temperature, and mechanical force intensity. These parameter adjustments allow dissociation to proceed while maintaining the fixed structural integrity of cells, ensuring both effective cell separation and preservation of morphological and biomolecular fidelity
2Stability of the object's composition
If conventional fixation methods are used, then tissue structure is preserved, but dissociation into single cells is compromised
Solution Approach 1:
The patent applies different fixation conditions to different tissue types and locations. By adjusting fixative concentration, penetration time, and temperature based on local tissue characteristics (density, vascularity, cell type), the method achieves optimal preservation of tissue structure in each region while maintaining compatibility with subsequent dissociation procedures, thus resolving the contradiction between structural preservation and dissociation efficiency
3Productivity
If rapid dissociation is performed, then processing time is reduced, but cell morphology and biomolecule integrity are compromised
Solution Approach 1:
The patent performs preliminary fixation that optimizes tissue permeability and structural stability before dissociation. This pre-treatment allows rapid dissociation to proceed without compromising morphology because the fixative has already established a stable framework that resists degradation during fast processing, thus resolving the contradiction between speed and precision
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 the faithful preservation of biomolecules and morphology in dissociated single cells, providing a reliable platform for analyzing the structure and composition of biological samples, reducing pre-analytical changes and improving diagnostic accuracy.
Implementation Method 1
fixing cells of the biological sample using a fixative wherein the fixative comprises a deep eutectic solvent
Implementation Method 2
dissociating the biological sample in the presence of the dissociation medium to obtain the single cells and/or groups of cells
Data Source
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AI summary
A method of processing a biological sample to obtain single cells and/or groups of cells comprises: fixing cells of the biological sample using a fixative; contacting the biological sample with a dissociation medium; dissociating the biological sample in the presence of the dissociation medium to obtain the single cells and/or groups of cells; wherein the fixative comprises a deep eutectic solvent. Fixing cells of the biological sample using a deep eutectic solvent facilitates dissociation of the biological sample into single cells, while at the same time inhibiting the degradation of biomolecules such as DNA and RNA in the cells. Also provided is an apparatus useful in the method, deep eutectic solvents and uses thereof, and a method of extracting cell nuclei.