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CyTOF uses antibodies with metal isotope labels to label cells and analyzes the composition of labels on each cell by time-of-flight mass spectrometry to conduct in-depth research on cell phenotype and function.

The working principle of CyTOF is analogous to that of FFC (Fluorescence Flow Cytometry), as both rely on binding target proteins with differently labeled antibodies for data generation. The only difference in CyTOF is that a pure heavy metal ion tag is applied in place of fluorochrome, and then it is subjected to mass spectrometry for data generation.

Single cell proteomics by mass cytometry.Fig. 1 Single cell proteomics by mass cytometry. (Prasad, 2019)

Advantages of CyTOF

  • A high-throughput tool that can measure more than 40 parameters per cell.
  • Low background signal.
  • Suitable for analysis of cell-surface and cytoplasmic proteins.
  • Applicable to fixed-cell analysis.
  • Mature data analytics.

Limitations of CyTOF

  • Typically requires a large sample size, owing to low recovery.
  • Not suitable for live-cell analysis.
  • Limited availability of metal-isotope-labeled antibodies.

The CyTOF Technical Workflow

  1. Tagging antibodies with stable, non-radioactive rare-earth-metal isotopes.
  2. Cells are immunostained with these tagged antibodies.
  3. Each labeled cell is then nebulized into a droplet and introduced into an argon plasma.
  4. The droplets containing single cells are then vaporized and the cellular components in the droplet are ionized.
  5. The ionized sample is passed into a mass filter, which allows only the heavy receptor ions through.
  6. These ions are then analyzed by a time-of-flight mass spectrometer.

CyTOF technical workflow.Fig. 2 CyTOF technical workflow. (Tracey, 2021)

Creative Biolabs has methodically developed every stage of the process, from sample preparation through data analysis, to assure high-quality study outcomes. If you have any requirements, please feel free to contact us for further communication about your project.

References

  1. Prasad, M.; et al. Chapter 14 – single-cell proteomics: technology and applications. Single-Cell Omics. 2019, 1: 283-318.
  2. Tracey, L.J.; et al. CyTOF: An emerging technology for single-cell proteomics in the mouse. Current Protocols. 2021, 1: e118.
! ! For Research Use Only. Not for diagnostic or therapeutic purposes.

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