Goto Toshiyuki, Nakano Masayuki, Danno Sally, Ueda Chie, Sakaue-Sawano Asako, Miyawaki Atsushi, Wrabel Anna, Nakahara Ichiro, Nishikata Takahito, Mizoguchi Akira, Tamura Yasuhisa, Mizuno Kei, Kataoka Yosky, Funabiki Kazuo
RIKEN Center for Biosystems Dynamics Research, Kobe, Hyogo 650-0047, Japan; Frontiers of Innovative Research in Science and Technology (FIRST), Konan University, Kobe, Hyogo 650-0047, Japan; Graduate School of Science, Technology and Innovation, Kobe University, Kobe, Hyogo 650-0047, Japan.
RIKEN Center for Biosystems Dynamics Research, Kobe, Hyogo 650-0047, Japan.
Cell Rep Methods. 2025 Jun 16;5(6):101056. doi: 10.1016/j.crmeth.2025.101056. Epub 2025 Jun 4.
The spatiotemporal heterogeneity in intratumor proliferative behavior of cancer cells deeply affects tumor environment characteristics and the efficacy of anticancer treatments. Thus, intravital imaging with unlimited imaging depth and cellular-level resolution is greatly desired. We developed an optical-fiber-bundle-based microendoscope with a genetically encoded fluorescent ubiquitination-based cell-cycle indicator (Fucci) system to achieve the intravital, periodic, and multicolor end-to-end imaging of the proliferative activity of cancer cells at a cellular-level resolution. This technique enabled the periodic visualization of spatiotemporal cellular responses, including cell-cycle arrest and resumption, and nuclear enlargement following the administration of anticancer drugs in living mice. It was suggested that proliferating cell ratio and nuclear enlargement in cancer cells at the surface region of tumor characterized by abundant vascular invasion contribute to aggressive tumor regrowth after chemotherapy. The application of this technique can accelerate innovation in cancer biology and therapeutics.
癌细胞肿瘤内增殖行为的时空异质性深刻影响肿瘤环境特征和抗癌治疗效果。因此，人们迫切需要具有无限成像深度和细胞水平分辨率的活体成像技术。我们开发了一种基于光纤束的微型内窥镜，结合基于基因编码荧光泛素化的细胞周期指示剂（Fucci）系统，以细胞水平分辨率实现对癌细胞增殖活性的活体、周期性和多色端到端成像。该技术能够对时空细胞反应进行周期性可视化，包括细胞周期停滞和恢复，以及在活体小鼠中给予抗癌药物后细胞核增大。研究表明，以丰富血管浸润为特征的肿瘤表面区域癌细胞的增殖细胞比例和细胞核增大有助于化疗后肿瘤的侵袭性再生。这项技术的应用可以加速癌症生物学和治疗学的创新。
