Selenium Maleimide Cell Barcoding Without Spectral Overlap
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Solution Overview
Problem
Current cell labeling and barcoding methods, such as mass cytometry, face limitations in parameterization due to spectral overlap and toxicity of reagents like ruthenium and osmium tetraoxide, and require pre-validation of antibodies for fixed epitope binding, restricting the number of parameters that can be determined and increasing toxicity risks.
Innovation Solution
Development of isotopically-pure selenium maleimide compounds that covalently react with cellular sulfhydryl groups, allowing for unique labeling of viable and fixed cells without spectral overlap and toxicity, enabling greater parameterization and deeper profiling capabilities.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If ruthenium and osmium tetraoxide are used for cell labeling, then cell barcoding capability is achieved, but toxicity increases and handling becomes dangerous
Solution Approach 1:
The patent changes the chemical identity of the labeling reagent from toxic ruthenium/osmium tetraoxide to less toxic selenium maleimide compounds, while maintaining the barcoding functionality through isotopic labeling of the selenium-containing compounds
Solution Approach 2:
The patent employs disposable selenium maleimide compounds that can be used for single-use cell barcoding applications, eliminating the need for expensive and hazardous metal reagents while achieving the same multiplexing capability
2Productivity
If fixed cells are barcoded and pooled for antibody staining, then sample pooling efficiency improves, but staining of chemokine receptors is excluded
Solution Approach 1:
The patent performs cell barcoding with selenium maleimide compounds before cell fixation and antibody staining, allowing subsequent staining of chemokine receptors on unfixed cells while maintaining the barcoding labels through the fixation process
Solution Approach 2:
The patent separates the barcoding function from the staining function, allowing different subsets of cells to be stained with different antibodies while all cells retain their barcoding labels, enabling flexible experimental designs
3Quantity of substance
If fluorescently tagged antibodies are used in flow cytometry, then parameter detection is enabled, but spectral overlap limits the number of parameters
Solution Approach 1:
The patent replaces the optical detection system of flow cytometry with mass cytometry detection, using selenium isotopes (76Se, 77Se, 78Se) that have distinct masses and no spectral overlap, enabling detection of many more parameters simultaneously
Solution Approach 2:
The patent changes the detection parameter from optical wavelength to mass-to-charge ratio, eliminating spectral overlap issues and enabling high-plex parameter detection with selenium-containing barcoding reagents
4Adaptability or versatility
If isotope-tagged antibodies are used to label surface markers, then cell barcoding is achieved, but the number of available parameters is reduced
Solution Approach 1:
The patent uses selenium maleimide compounds that can serve both as barcoding reagents and as universal labels for any cell type, freeing up antibody-based parameters for specific marker detection without competing for the same isotopic labels
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
The selenium maleimide compounds effectively label cells with minimal channel spill, allowing for greater parameterization and deeper profiling without the need for antibodies, reducing toxicity and enabling uninterrupted instrument runs and improved data consistency.
Implementation Method 1
isotopically-pure selenium maleimide compounds that covalently react with cellular sulfhydryl groups
Data Source
AI summary
The present disclosure relates to selenium containing compounds useful as cellular labeling and barcoding reagents, such as isotopically-pure selenium maleimide compounds and the use thereof.