Related Experiment Video
Updated: Sep 22, 2025
Sample Preparation for Mass Cytometry Analysis
Published on: April 29, 2017
Full spectrum flow cytometry and mass cytometry: A 32-marker panel comparison
Maria C Jaimes1, Michael Leipold2, Geoffrey Kraker3
1Applications, Cytek Biosciences, Inc, Fremont, California, USA.
Insights
Full spectrum flow cytometry (FSFC) offers a powerful alternative to mass cytometry (MC) for high-dimensional immune monitoring. This study demonstrates highly comparable results between FSFC and MC using a 32-marker immune panel.
Area of Science:
- Immunology
- Biotechnology
- Single-cell analysis
Background:
- High-dimensional single-cell data is crucial for understanding immune system complexity in health and disease.
- Mass cytometry (MC) has been the primary technology for assays exceeding 29 markers.
- Full spectrum flow cytometry (FSFC) now enables analysis beyond the traditional fluorescence limits.
Purpose of the Study:
- To directly compare the performance of MC and FSFC platforms.
- To evaluate the comparability of results using an established 32-marker immune panel.
- To assess the utility of FSFC in immune subpopulation identification.
Main Methods:
- Comparison of five patient samples using both MC and FSFC.
- Utilized an established 32-marker immune panel from the Stanford Human Immune Monitoring Center (HIMC).
- Employed split samples and identical antibody panels for direct technology comparison.
Main Results:
- Demonstrated highly comparable results between MC and FSFC across multiple data analysis approaches.
- Confirmed the ability of both platforms to identify all expected immune subpopulations with the 32-marker panel.
- Validated FSFC as a viable alternative to MC for complex immune monitoring.
Conclusions:
- FSFC provides comparable performance to MC for high-dimensional immune monitoring.
- FSFC expands the possibilities for cytometric assays beyond 29 parameters.
- This comparison supports the adoption of FSFC for detailed immune cell analysis.
Abstract:
High-dimensional single-cell data has become an important tool in unraveling the complexity of the immune system and its involvement in homeostasis and a large array of pathologies. As technological tools are developed, researchers are adopting them to answer increasingly complex biological questions. Up until recently, mass cytometry (MC) has been the main technology employed in cytometric assays requiring more than 29 markers. Recently, however, with the introduction of full spectrum flow cytometry (FSFC), it has become possible to break the fluorescence barrier and go beyond 29 fluorescent parameters. In this study, in collaboration with the Stanford Human Immune Monitoring Center (HIMC), we compared five patient samples using an established immune panel developed by the HIMC using their MC platform. Using split samples and the same antibody panel, we were able to demonstrate highly comparable results between the two technologies using multiple data analysis approaches. We report here a direct comparison of two technology platforms (MC and FSFC) using a 32-marker flow cytometric immune monitoring panel that can identify all the previously described and anticipated immune subpopulations defined by this panel.
