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Mass Cytometry: Protocol for Daily Tuning and Running Cell Samples on a CyTOF Mass Cytometer
Published on: November 2, 2012
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Tantalum Oxide Nanoparticle-Based Mass Tag for Mass Cytometry
Yefeng Zhang1, Nick Zabinyakov2, Daniel Majonis2
1Department of Chemistry, University of Toronto, 80 St. George Street, Toronto, Ontario M5S 3H6, Canada.
Analytical Chemistry
|April 3, 2020
Summary
Researchers developed novel tantalum oxide nanoparticle (NP)-based mass tags for mass cytometry (MC). This advancement enables higher dimensional single-cell analysis, expanding the potential of MC immunoassays.
Area of Science:
- Biotechnology
- Analytical Chemistry
- Immunology
Background:
- Mass cytometry (MC) is a high-dimensional single-cell analysis technique using metal-tagged antibodies.
- Current MC methods can measure over 40 parameters, with potential for up to 135 parameters.
- Development of new elemental mass tags is crucial for expanding MC capabilities.
Purpose of the Study:
- To develop and characterize tantalum oxide nanoparticle (NP)-based mass tags for MC immunoassays.
- To demonstrate the utility of these novel NP-based mass tags in high-dimensional single-cell analysis.
- To integrate these new reagents into existing MC workflows.
Main Methods:
- Synthesized uniform-sized amine-functionalized tantalum oxide NPs using a reverse microemulsion method.
- Modified NPs with azide groups and conjugated them to dibenzocyclooctyne (DBCO)-functionalized antibodies (Abs).
- Fabricated and tested three specific Ab-NP conjugates (TaO2-PEG2-goat antimouse, TaO2-PEG2-CD25, TaO2-PEG2-CD196) in MC immunoassays.
Main Results:
- Successfully synthesized and functionalized tantalum oxide NPs for antibody conjugation.
- Demonstrated effective detection of CD20 biomarkers on Ramos cells using a TaO2-PEG2-goat antimouse conjugate.
- Showcased successful integration of TaO2-PEG2-CD25 and TaO2-PEG2-CD196 conjugates into commercial panels for immune profiling.
Conclusions:
- Tantalum oxide NPs represent a viable new mass tag for MC immunoassays.
- This NP-based approach enhances the multiplexing capacity of MC.
- The developed conjugates facilitate high-dimensional single-cell immune profiling.
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