Mass Cytometry

CyTOF XT Mass Cytometer

Cytometry and Time-Of-Flight Mass Spectrometry

The CyTOF XT mass cytometer analyzes individual cells labeled with stable heavy metal isotypes using Time-of-Flight Inductively Coupled Plasma mass spectrometry. The CyTOF XT can simultaneously resolve 30-40 multiple elemental probes on a per-cell basis without the need for spillover compensation between channels. 

Essential Steps for Success

• ​Environmental contaminants contain metals that will be detected by the CyTOF XT. Common detergents frequently contain Barium. Avoid using plastic or glassware that has ever been washed with detergent.
• Use only reagents that have been verified to be metal-free.
• Filter all home-made reagents using 0.2 um filters.
• Use only ultrapure Milli-Q water for reagents.
  

When compared to standard flow cytometry, cell loss is greater in CyTOF XT experiments due to the multiple wash steps required in staining, resuspension of fixed cells in water prior to introduction into the system, and loss of cells within the mass cytometer. Plan on staining more cells than you would need for a traditional flow cytometry experiment.

Mass Cytometry Staining Protocols

Antibody Bank

To minimize the costs associated with purchasing large amounts of antibodies for mass cytometry sample staining, we have established a shared antibody bank. The bank is available for all AMC investigators. MaxPar antibodies may be purchased in small volumes from the bank. If you need access to additional MaxPar antibodies, please contact us at cc.flowcyto@ucdenver.edu.

To take advantage of this offering, please submit an order request through our CyTOF Helios Antibody Bank. Please note that orders may take up to 5 days to complete.

Helpful Resources

• Standard BioTools: The company that sells all mass cytometry instruments and isotopically-labelled antibodies
• Standard BioTools antibody catalog
• Commerically-available CyTOF antibody panels
• Stanford Cyto Forum: A compendium of questions and answers on using Mass Cytometry. A great place to investigate if you have questions about mass cytometry. 
• Nolan Lab​