Rapid native single cell mass spectrometry
Inventors
Assignees
Publication Number
US-12050221-B2
Publication Date
2024-07-30
Expiration Date
2039-11-15
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Abstract
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.
Core Innovation
The invention provides a method and system for analyzing single cells by mass spectrometry, utilizing a process whereby single cells are isolated from a sample in liquid medium, ejected individually, and captured in a specially designed capture probe containing a flowing capture probe solvent. Upon capture, each cell is lysed by a lysis inducer inside the probe, dispersing the cellular components into the medium, after which these components are transported to a mass spectrometer for analysis. Each lysed cell’s components are spatially and temporally separated from those of other cells to ensure individual analysis.
This invention addresses the analytical challenge of detecting metabolites and other molecular species at the single-cell level without extensive sample preparation or perturbation of the cell's natural state. Existing technologies either require prior knowledge of analytes, suffer from low throughput, or necessitate significant cell preparation that may alter native cell chemistry. The invention circumvents these limitations by enabling untargeted, high-throughput, quantitative mass spectrometric analysis of individual, unmodified cells directly from their native environment.
Lysis of cells within the capture probe can be induced by various means, including use of an appropriate solvent with lower partial pressure, chemical lysing agents, mechanically induced liquid vortices, application of voltage, or acoustic waves. The system is structured such that the liquid vortex is formed by a coaxial probe, allowing on-demand and precise ejection and capture of single cells, reducing the risk of sample carryover and preserving the integrity of each cell’s chemical profile through minimal dilution and rapid analysis.
Claims Coverage
There are two independent claims, each detailing a method and a system, both centered on the use of a coaxial probe capture system and spatial/temporal separation of single-cell content during mass spectrometric analysis.
Method for single cell mass spectrometry with coaxial probe spatial/temporal separation
This method involves: - Providing a plurality of cells in a liquid medium and placing them in a single cell isolation and ejection system. - Releasing individual cells contained in liquid droplets from the system. - Capturing each single cell with its medium in a capture probe containing a flowing capture probe solvent. - The probe is a coaxial probe where solvent flows from a supply path, overflows a common open end, and enters a coaxial exhaust flow path. - Lysing the cell in the capture probe by a lysis inducer, dispersing cell components into the medium. - Transporting lysed single cell components to a mass spectrometer, ensuring spatial and temporal separation from contents of other single cells entering the spectrometer. - Conducting mass spectrometry on the separated lysed single cell components.
System for single cell mass spectrometry with coaxial probe and lysis in capture probe
This system includes: - A single cell isolation and ejection system with a storage container for plurality of cells in a liquid medium. - A capture probe comprising at least one solvent supply conduit for flowing a capture probe solvent and a capture probe exhaust conduit for receiving single cell droplets and conducting lysed cell components to a mass spectrometer. - The capture probe is a coaxial probe wherein the solvent flows from the solvent supply conduit, overflows a common open end of the coaxial probe, and enters the probe exhaust conduit. - The probe includes a lysis inducer to lyse cells once captured in the probe before transfer to the mass spectrometer.
The inventive features define both a unique method and a system for single cell analysis by mass spectrometry, emphasizing coaxial probe design, in-probe lysis with various inducers, and spatial/temporal separation of individual cell components during analysis.
Stated Advantages
Enables rapid, untargeted, and quantitative mass spectrometric analysis of single cells in their native medium.
Requires no extensive sample preparation or molecular labeling, preserving cells in their natural state until analysis.
Provides high sensitivity and the ability to detect and quantify a wide range of molecular species from small metabolites to large biopolymers.
High throughput is achievable, allowing for statistically robust analysis of single cell populations or subpopulations.
Eliminates signal carryover and cross-contamination between individual cell analyses through continuous solvent flow in a self-cleaning probe.
Compatible with nearly any cell suspension, including mammalian cells, without the need for modification.
Enables real-time chemical classification and differentiation of single cells using mass spectral data.
Documented Applications
Single cell lipid and metabolite profiling of microalgae (Chlamydomonas reinhardtii and Euglena gracilis) and mammalian cells (HeLa cells) for distinction and quantification of chemical differences between cell types or growth conditions.
Untargeted and targeted chemical analysis of single cells directly from cell culture media with minimal sample preparation.
Real-time classification and differentiation of single cells in mixtures, such as distinguishing cell types in biological samples or potentially in clinical samples (e.g., blood, tumors).
Quantitative analysis of lipid distributions in single cells to study effects of environmental changes, such as nutrient deprivation.
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