Single-Cell Mass Spectrometry Flow Cytometry Analysis

With the development of modern biology, the term 'average' can no longer satisfy our research needs. We need to further understand the differences between cells, which has led to the emergence of various single-cell research technologies. Mass Cytometry is a flow technology that uses mass spectrometry to perform multi-parameter detection on single cells. It combines the high-speed analysis capabilities of traditional flow cytometry with the high-resolution capacity of mass spectrometry. Both single-cell sequencing and mass cytometry are currently very powerful technologies, and more and more high-profile publications are combining the two, without saying one is better than the other. The difference lies in mass cytometry detecting proteins, allowing the specification of proteins to be detected, avoiding missing low-abundance yet important proteins. Additionally, mass cytometry technology, with its fairly high throughput, can detect a large number of cells, making it a good complement to single-cell sequencing. Becton Dickinson Biosciences' mass cytometry technology uses metal element labels (usually metal-labeled specific antibodies) to mark molecules on the cell surface and interior, then separates individual cells using flow cytometry principles, analyzes the atomic mass spectrum of individual cells using inductively coupled plasma mass spectrometry (ICP-MS), and finally converts the atomic mass spectrum data into expression levels of signaling molecules on the cell surface and inside the cell.

McCarthy, R. L. et al. J. Vis. Exp. 2017.

Mass Cytometry Technology

Advantages of Mass Cytometry Technology:

1. Advanced technology, filling the technological gap

Mass cytometry adopts metal-labeled antibody technology, avoiding the problem of few and easily interfering traditional fluorescence channels. It can simultaneously characterize multiple indicators at the single-cell level, with Becton Dickinson Biosciences capable of detecting 51 target proteins at the same time.

2. Large analysis quantity, relatively low cost

Single-cell RNAseq is limited by cost and other factors, with the total number of cells analyzed across all samples generally around 2x10^4. In contrast, mass cytometry can analyze at least 10^5 cells in a single sample, achieving a magnitude increase without costing more than single-cell RNAseq.

3. Great application prospects

a) Mass cytometry results can reveal changes in cell subpopulations, offering great research prospects in clinical diagnostics and disease mechanism studies;
b) Combining metal labeling technology with other technologies can lead to new application directions. Beyond conventional proteins, mass cytometry can also be used for protein post-translational modifications;
c) It can detect cell viability, cell size, mRNA transcript expression levels, DNA synthesis rate, and protease activity, etc.

Mass Cytometry Workflow
Becton Dickinson Biosciences uses the Fluidigm mass cytometry system for sample analysis

 

Mass Cytometry Data Analysis

Applications of Mass Cytometry Technology
Mass cytometry's functional analyses can be divided into 12 categories:
(A) Phenotypic characterization
(B) Intracellular cytokine determination
(C) Characterization of intracellular signaling states
(D) Measurement of cell volume and size
(E) Identification of cell viability
(F) Cell cycle identification
(G) Proliferation tracking
(H) Receptor occupancy measurement
(I) Tetramer-based antigen-specific T cell screening
(J) Chromatin modification analysis
(K) Co-detection of RNA and protein
(L) Imaging mass cytometry

Mass cytometry has widespread applications in disease research and treatment, taking the study of novel coronaviruses as an example:

 

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