Fig. 1.

Workflow for imaging mass cytometry on ocular FFPE tissue. Ocular tissue is removed during surgery, followed by fixation, paraffin embedding and sectioning using a microtome. Sections are stained with metal-labeled antibodies. Within the Hyperion Imaging Mass Cytometry system a laser ablates the tissue point by point (spot size 1 µm²). Laser ablation and subsequent vaporization creates particle clouds, which are transferred into the mass cytometer by a stream of inert gas. The mass cytometer measures the composition of each particle cloud and reconstructs an image for each stained marker based on the measurement. The resulting images can be processed and analyzed using bioinformatics tools. Cell segmentation allows the analysis of marker expression in each individual cell, providing the data for bioinformatic single-cell analysis