Single Cell Mass Cytometry Analysis

With the advancement of modern biology, the term "average" no longer satisfies our research needs. We need to delve deeper into the differences between individual cells, leading to the emergence of various single-cell research technologies. Single-cell mass cytometry employs mass spectrometry to perform multi-parameter detection on individual cells, combining the high-throughput analysis capabilities of traditional flow cytometry with the high resolution of mass spectrometry detection. Both single-cell sequencing and mass cytometry are currently very powerful techniques, and more and more high-impact publications are using a combination of the two, with no superiority between them. The difference lies in that mass cytometry targets proteins, allowing for the detection of specified proteins, avoiding the omission of low-abundance yet important proteins, and due to its high throughput, it can analyze a large number of cells, complementing single-cell sequencing well. MtoZ Biolabs' single-cell mass cytometry technology utilizes metal element markers (typically metal-labeled specific antibodies) to label molecules on the cell surface and inside, then separates individual cells using flow cytometry principles, followed by analysis of the atomic mass spectrum of each cell using inductively coupled plasma mass spectrometry (ICP-MS), and finally converting the atomic mass spectrum data into the expression levels of signal molecules on the cell surface and inside.

 



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

Figure 1. Single-Cell Mass Cytometry Technology

 

Service Advantages

1. Advanced Technology, Filling Technical Voids 

Single-cell mass cytometry uses metal-labeled antibody technology, which avoids the issues of limited and mutually interfering fluorescent channels typical of traditional flow cytometry. It enables the simultaneous characterization of multiple markers at the single-cell level, with MtoZ Biolabs capable of simultaneously detecting 51 target proteins.

 

2. Large Analysis Volume, Relatively Low Cost 

The number of cells analyzed across all samples in single-cell RNA sequencing is generally limited to about 20,000 due to cost and other factors, whereas mass cytometry technology can analyze at least 100,000 cells in a single sample, achieving a significant increase in volume and maintaining a cost not higher than that of single-cell RNA sequencing.

 

3. Great Application Prospects

① Mass cytometry results can reveal changes in cell subgroups, offering significant research potential in clinical diagnostics and disease mechanism studies.

② Combining metal labeling technology with other techniques opens new application directions. Beyond conventional proteins, mass cytometry can also be used for post-translational modifications of proteins.

③ It can measure cell viability, cell size, mRNA transcript expression, DNA synthesis rate, and protease activity.

 

Analysis Workflow

Uses the Fluidigm mass cytometry system for sample analysis

1. Experimental Design

With the use of the Maxpar® Control Panel Designer, it is easy to construct control panels comprising up to 50 metal-tagged antibodies and reagents. You can also enhance data quality by duplicating your samples with the addition of barcodes or utilize pre-made control panels from Fluidiam!

 

2. Sample Preparation

The CyTOF system can acquire highly multiplexed single-cell data from suspended cell samples. Standard protocols for preparing single-cell suspensions from solid or liquid tissues and tumors are also applicable for CyTOF sample preparation.

 

3. Data Acquisition

The CyTOF XTᵀᴹ system features complete automation of particle resuspension, calibration bead addition, and batch sample acquisition from an auto-sampler conveyor. The Heliosᵀᴹ system offers data quality standards comparable to the CyTOF XT, but requires manual resuspension and loading of samples.

 

4. Data Analysis

The CyTOF systems can generate standard FCS 3.0 (Helios) or 3.1 (CyTOF XT) data files, which can be analyzed using any compatible software, including FCS Expressᵀᴹ and Maxpar Pathsetterᵀᴹ.

 

Applications

Functional analysis that can be performed with mass cytometry can be divided into 12 categories:

1. Phenotypic Characterization

2. Intracellular Cytokine Determination 

3. Intracellular Signaling Status Characterization 

4. Cell Volume and Size Measurement 

5. Cell Vitality Identification

6. Cell Cycle Identification 

7. Proliferation Tracing 

8. Receptor Occupancy Determination 

9. T-cell Screening Based on Tetramers 

10. Chromatin Modification Analysis 

11. Co-detection of RNA and Proteins 

12. Imaging Mass Cytometry

 

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