Research Techniques Made Simple: Experimental Methodology for Imaging Mass Cytometry.

Technological advances in flow cytometry and the development of mass cytometry by time-of-flight (CyTOF) have led to progressive increases in the number of proteins and biochemical processes that can be simultaneously measured. The most recent development of these platforms, imaging mass cytometry (IMC), allows for the visualization of up to 40 unique cellular markers and also employs rare metal isotopes conjugated to antibodies. However, IMC also adds the important benefit of preserving two-dimensional (2D) tissue architecture; this is accomplished by staining in situ and direct tissue vaporization followed by generation of a 2D spectral reconstruction using CyTOF-captured events. We review the experimental methodology for IMC that enables high-resolution multilayer images depicting protein expression, cellular localization, and interaction in situ in dermatology research.