Tanaka Koji, Toiyama Yuji, Okugawa Yoshinaga, Okigami Masato, Inoue Yasuhiro, Uchida Keiichi, Araki Toshimitsu, Mohri Yasuhiko, Mizoguchi Akira, Kusunoki Masato
Department of Gastrointestinal and Pediatric Surgery, Mie University Graduate School of Medicine 2-174 Edobashi, Tsu, Mie 514-8507, Japan.
Department of Neural Regeneration and Cell Communication, Mie University Graduate School of Medicine 2-174 Edobashi, Tsu, Mie 514-8507, Japan.
Am J Transl Res. 2014 May 15;6(3):179-87. eCollection 2014.
Intravital (in vivo) microscopy using fluorescently-tagged proteins is a valuable tool for imaging the expression of a specific protein, its subcellular location and the dynamics of specific cell populations in living animals. Recently, multiphoton microscopy including two-photon laser scanning microscopy (TPLSM) has been used in the field of tumor biology due to its ability to image target organs at higher magnification and at deeper depths from the tissue surface for longer time periods. We developed a method of in vivo real-time imaging for tumor metastasis using TPLSM with an organ stabilizing system, which allow us to observe not only a single tumor cell and its microenvironment for a long time, but also to observe the same organ of the same mouse at multiple time points in preclinical models. Here, we presented in vivo real-time images of 1) tumor cell arrest, 2) tumor cell-platelet interaction, 3) tumor cell-leukocyte interaction, and 4) metastatic colonization at the secondary organs as representative steps of metastatic process of experimental liver metastasis models using TPLSM.
使用荧光标记蛋白的活体（体内）显微镜检查是一种用于成像特定蛋白表达、其亚细胞定位以及活体动物中特定细胞群体动态的重要工具。最近，包括双光子激光扫描显微镜（TPLSM）在内的多光子显微镜已被应用于肿瘤生物学领域，因为它能够在更高放大倍数下对目标器官进行成像，并能从组织表面更深的深度进行更长时间的成像。我们开发了一种使用带有器官稳定系统的TPLSM进行肿瘤转移体内实时成像的方法，这使我们不仅能够长时间观察单个肿瘤细胞及其微环境，还能在临床前模型中在多个时间点观察同一只小鼠的同一器官。在此，我们展示了使用TPLSM对实验性肝转移模型转移过程的代表性步骤进行的体内实时成像，包括1）肿瘤细胞停滞、2）肿瘤细胞与血小板相互作用、3）肿瘤细胞与白细胞相互作用以及4）在次级器官的转移定植。
