![]() Additionally, we outline novel findings obtained by this new technique. In this review, we focus on the emerging role of intravital imaging in gastrointestinal research and describe how to observe the intestines and immune cells using intravital microscopy. In recent years, the application of intravital microscopy in gastrointestinal research has provided novel insights into the mechanisms of intestine-specific events including innate and adaptive immunities. Additionally, understanding the immune responses involved in gastrointestinal tumors and tissue repair is becoming increasingly important for the elucidation of disease mechanisms that have been unknown. Understanding the in vivo behavior of these immune cells and their function in gastrointestinal inflammation, including colitis, inflammatory bowel disease, ischemia–reperfusion injury, and neutrophil extracellular traps, is critical for gastrointestinal research to proceed to. To date, many kinds of immune cells have been identified, but their precise roles in intestinal immunity remain unclear. In an era of single-cell transcriptomics and data-driven research, 2P-IVM will remain a key advantage in delivering the missing spatiotemporal context in the field of cancer research. We also discuss the potential application of red-shifted indicators along with optogenetic tools to 2P-IVM. ![]() In this review, we summarize the current state of 2P-IVM with fluorescent indicators of signal transduction to reveal the cross-talks between cancer cells and surrounding cells including both immune and non-immune cells. ![]() technique for observing intercellular cross-talks in situ, unraveling cellular and molecular mechanisms in the context of their spatiotemporal dynamics. ![]() Today, intravital microscopy with two-photon excitation microscopes (2P-IVM) is the mainstay. The inflammatory responses in the tumor microenvironment are shaped by complex sequences of dynamic intercellular cross-talks among diverse types of cells, and recapitulation of these dynamic events in vitro has yet to be achieved. Inflammation can contribute to the development and progression of cancer. ![]()
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