Fig. 2

2P-IVM with fluorescent indicators to reveal the cross-talks between tumor cells and surrounding cells in the tumor microenvironment including both immune and non-immune cells. Indirect cross-talks between A tumor cells and non-immune cells, B tumor cells and immune cells, and C direct cross-talks between tumor cells and immune cells can be visualized by using 2P-IVM with fluorescent indicators (gray cells: tumor cells; magenta cells: tumor cells receiving signals via signaling molecules, TXA₂ or IFN-γ; yellow cells: T cells or NK cells). Melanoma cells expressing GCaMP6s were imaged using 2P-IVM (A, bottom). The image represents the maximum intensity projection for 10 min. Images were adapted courtesy of Konishi et al. [36]. Schematic representation of the fluorescent indicator for IFN-γ (B, bottom). Melanoma cells expressing this fluorescent indicator were imaged using 2P-IVM. Images represent the mCherry and turquoise-GL-NLS of parental cells. Images were adapted by courtesy of Tanaka et al. [38]. Melanoma cells expressing GCaMP6s were imaged in the lung using 2P-IVM (C, bottom). Top, FRET/CFP ratio of an NK cell is shown. Melanoma cell is shown in white. GCaMP6s intensity is displayed in pseudo-color (bottom). The NK cell is shown in white. The images were adapted by courtesy of Ichise et al. [39]