Single-Cell Mass Spectrometry With Liquid Vortex Capture
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Solution Overview
Problem
Current mass spectrometry techniques face challenges in rapidly and routinely acquiring quantitative molecular information from single cells due to low sampling throughput and the need for extensive sample preparation, which can alter cellular chemistry and limit the analysis to specific molecules, making it difficult to detect the complex and dynamic metabolites within cells.
Innovation Solution
A method involving a single cell isolation and ejection system that uses a capture probe with a flowing solvent to lyse cells and transport components to a mass spectrometer, allowing for spatial and temporal separation of single cell components, enabling high-throughput analysis without prior sample preparation and maintaining cells in their native state until analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If vacuum-based MS techniques (SIMS, MALDI, LDI) are used to achieve high sensitivity, then detection sensitivity is improved, but sample preparation complexity increases and cells must be fixed and dehydrated which alters their natural state
Solution Approach 1:
The patent replaces vacuum-based mechanical ionization systems with an ambient pressure liquid vortex capture system. Cells are lysed in a liquid vortex at atmospheric pressure, and analytes are transferred to the mass spectrometer without requiring vacuum-based sample preparation steps like fixation and dehydration. This substitution eliminates the need for complex sample preparation while maintaining detection sensitivity.
Solution Approach 2:
The patent introduces a liquid vortex as an intermediary medium between the cell sample and the mass spectrometer. The liquid vortex captures cell lysates and transports them to the MS interface, serving as a bridge that eliminates the need for direct vacuum-based analysis. This intermediary approach allows cells to be analyzed in their native state without extensive preparation.
2Ease of operation
If manually guided probes (pipettes, optical fibers, capillaries) are used for ambient MS to measure cells in situ, then cells remain in native state, but sampling throughput decreases to less than 1 cell/s
Solution Approach 1:
The patent uses a liquid vortex hydraulic system to rapidly capture and process multiple cells per second. The flowing liquid vortex creates a dynamic capture zone that can accommodate high-throughput cell delivery while maintaining ambient pressure conditions. This hydraulic approach replaces manual probing with an automated fluid-based system that preserves native state while achieving high throughput.
Solution Approach 2:
The patent performs preliminary cell lysis in the liquid vortex before mass spectrometry analysis. By pre-lysing cells in the capture probe, the system eliminates the need for slow manual manipulation during analysis. This preliminary action allows rapid processing of multiple cells while maintaining their chemical integrity until the moment of analysis.
3Difficulty of detecting and measuring
If laser ablation techniques are used to dissect single cells for MS analysis, then tissue sampling is improved, but sampling throughput and sensitivity remain low for cell suspension analysis
Solution Approach 1:
The patent replaces laser ablation mechanical disruption with a liquid vortex-based lysis approach. Instead of using high-energy laser pulses to dissect cells, the system uses the mechanical and chemical action of a flowing liquid vortex to lyse cells and release analytes. This substitution improves throughput by eliminating the slow, sequential nature of laser ablation while maintaining effective cell disruption.
4Productivity
If flow cytometry is used to achieve high throughput (thousands of cells/s), then sampling speed is improved, but molecular analysis is limited to fluorescence-based probes targeting only a dozen molecules simultaneously
Solution Approach 1:
The patent creates a universal analysis platform that combines high-throughput liquid vortex capture with untargeted mass spectrometry. Unlike flow cytometry's probe-specific approach, this system can detect any ionizable molecule in the cell lysate without requiring prior knowledge of target molecules. The liquid vortex capture mechanism is universal and can handle diverse cell types, while the MS detector provides universal molecular coverage across thousands of metabolites simultaneously.
Solution Approach 2:
The patent uses mass spectrometry to create a comprehensive molecular copy of the cell's chemical state. Instead of using fluorescence probes that only detect specific labeled molecules, the MS system captures and identifies a broad range of metabolites based on their mass-to-charge ratio. This molecular copying approach provides versatile, untargeted analysis while maintaining high throughput through the liquid vortex interface.
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 approach enables rapid, untargeted, and quantitative analysis of single cell chemistry with high sensitivity, allowing for the differentiation of cellular populations and subpopulations based on metabolite profiles, and can process up to 100 cells per second, significantly improving the throughput and accuracy of single cell mass spectrometry.
Implementation Method 1
a liquid vortex formed by the flowing capture probe solvent
Implementation Method 2
Mass spectrometry can then be conducted on the lysed single cell components. The lysed single cell components entering the mass spectrometer are spatially and temporally separated from any dispersed components of another single cell
Data Source
AI summary
A method for analyzing single cells by mass spectrometry includes the steps of providing a plurality of cells in a liquid medium and placing the cells and liquid medium in a single cell isolation and ejection system. Liquid medium containing a single cell is released from the single cell isolation and ejection system. The liquid medium and single cell are captured in a capture probe containing a flowing capture probe solvent. The cell is lysed by a lysis inducer in the capture probe to disperse single cell components into the medium. The lysed single cell components are transported to a mass spectrometer, where the lysed single cell components entering the mass spectrometer are spatially and temporally separated from any dispersed components of another single cell from the sample entering the mass spectrometer. Mass spectrometry is conducted on the lysed single-cell components. A system for analyzing single cells by mass spectrometry is also disclosed.