Reliable and sensitive detection of rare events
Reliable and sensitive detection is critical in rare event analysis. ImmunoScape uses proven tetramer technology as the foundation for single-cell, high-dimensional, rare-event detection.
In a 2009 Nature Methods paper, published by Newell et al., the simultaneous use of large numbers of different peptide-MHC Class I (pMHC-I) tetramers on cells using standard flow cytometry and reagents was demonstrated.
Due to the limitations of flow cytometry, including crosstalk between the limited repertoire of then available fluorophores, the tetramer multiplexing method was extended to a mass cytometry platform (CyTOF®) to enable broader and deeper profiling of antigen-specific T cells.
Using ten metal labels with combinatorial MHC-I tetramer staining in unique combinations of three metal labels per MHC multimer, 120 antigen specificities can be screened per sample, with a substantial number of labels, i.e. 30 or more, still available for parallel phenotyping and functional analysis of the T cells.
Tetramers enable analysis of the characteristics of the antigen-specific T cells ex vivo without perturbing the cells or changing their phenotypes. Cells are stained for a very short period of time at low temperature so they retain their phenotypes and can even be detected in a completely dysfunctional state.