Establishment of an Academic Tissue Microarray Platform as a Tool for Soft Tissue Sarcoma Research

Figure 3

Example of marker expression using multiplex immunofluorescence (MILAN), on tissue microarray from tissue cores of alveolar soft part sarcoma. Representative fluorescence immunostainings of tissue cores and different cycles of the identical core showed the stability of tissue microarray throughout experiments. Images were digitally scanned under 200-fold magnification. The blue color showed DAPI staining and represented nucleated cells. Membrane expression of programmed cell death ligand 1 (red) was detected in a small proportion of tumor cells in the first cycle (left column) but no expression for programmed cell death 1 (green) was observed. In the middle column, pink color in merged images indicated nuclear localization of transcription factor E3 (red) but no green fluorescence for cytotoxic T lymphocyte-associated protein 4 was detected. In the sixth cycle (right column), membrane expression of major histocompatibility complex I/II and their coexpression were demonstrated in red, green, and yellow, respectively.