炎症可加速癌症的发展和进展。 肿瘤微环境中的炎性反应受到不同类型细胞之间复杂的动态细胞间相互作用的影响，在体内重现这些动态事件尚未实现。 如今，具有双光子激发显微镜（2P-IVM）的体内显微镜是观察体内细胞间相互作用、揭示其在时空动态下的细胞和分子机制的主要技术。 在本综述中，我们总结了具有信号转导荧光指示剂的2P-IVM的当前状态，以揭示癌细胞和周围细胞（包括免疫和非免疫细胞）之间的相互作用。 我们还讨论了红移指示剂与光遗传工具在2P-IVM中的潜在应用。 在单细胞转录组学和数据驱动研究的时代，2P-IVM仍将是在癌症研究领域中提供缺失的时空背景的关键优势。 © 2023年作者。

Inflammation can contribute to the development and progression of cancer. 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. Today, intravital microscopy with two-photon excitation microscopes (2P-IVM) is the mainstay technique for observing intercellular cross-talks in situ, unraveling cellular and molecular mechanisms in the context of their spatiotemporal dynamics. 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. We also discuss the potential application of red-shifted indicators along with optogenetic tools to 2P-IVM. 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.© 2023. The Author(s).