CyTOF

Mass cytometry is a single-cell proteomic technology that allows for the analysis of 40+ cellular parameters using antibodies tagged with heavy metal isotopes. The readout of this technology is based on mass spectrometry, resulting in signals for each individual cell that correlate to the expression of the target proteins.

The data output is similar to flow cytometry, but with the advantages of a higher number of concurrent parameters and the absence of fluorescence compensation.  This allows for a greater depth and breadth of cellular analysis within a given sample. In addition, palladium isotope barcoding may be used to label up to 20 separate samples that can then be pooled into a single tube for staining and analysis.

The CIMACs will offer CyTOF analysis for PBMC samples using a standardized panel of over 30 markers, delineating immune populations as well as their activation status and checkpoint marker expression. Current panel design is under validation across CIMAC centers and includes 14 immunological core markers (Table 1). Additional markers will be added after discussion among CIMAC centers.

Mt Sinai: Core Lyophilized Panel

TargetClone
CD19HIB19
CD45RAHI100
CD4RPA-T4
CD8aRPA-T8
CD163G8
CD1cL161
CD1236H6
CD66bG10F5
CD27O323
CD14M5E2
CD56B159
CD3UCHT1
CD38HB-7
HLADRL243
Table 1: Basic Initial CIMAC CyTOF Panel.

Final Panel Design with additional markers (up to 40) under optimization.

CyTOF panels are available at Stanford, Dana-Farber, and Mount Sinai CIMAC sites, and will be available at MD Anderson CIMAC soon.