Intravital dual-colored visualization of colorectal liver metastasis in living mice using two photon laser scanning microscopy
Intravital dual-colored visualization of colorectal liver metastasis in living mice using two photon laser scanning microscopy
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DOI：
10.1002/jemt.21059
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发表时间：
2012-03-01
影响因子：
2.5
通讯作者：
Kusunoki, Masato
中科院分区：
文献类型：
--
作者：
Tanaka, Koji;Morimoto, Yuhki;Kusunoki, Masato
癌症生物学的一个主要挑战是以高光学分辨率在体内实时可视化次级器官中转移过程的动态。在这里，我们提出了活体，双色成像的肝转移形成从一个单一的癌细胞转移的殖民地在活的小鼠的活肝使用双光子激光扫描显微镜（TPLSM）。将表达红色荧光蛋白的鼠（SL 4）或人（HT 29）结肠直肠癌细胞系接种到表达绿色荧光蛋白的小鼠的脾脏中。活体TPLSM通过活体小鼠肝叶的外置和固定来进行。这被重复多次，用于同一小鼠的长期成像。活小鼠的活肝脏中的活癌细胞在超过600倍的放大倍数下活体内可视化。注射后2小时，单个癌细胞被阻滞在肝窦内。观察到癌细胞周围的血小板聚集，表明肿瘤细胞诱导的血小板聚集现象。癌细胞从肝窦渗出到Disse间隙。观察到癌细胞周围的枯否细胞突起，表明枯否细胞似乎吞噬癌细胞。SL 4细胞形成肝转移集落，具有广泛的间质反应。HT 29细胞的肝转移被观察为一簇微转移结节。使用活体TLPSM的癌症转移的高分辨率，双色，实时可视化可以帮助理解在活体动物的活体器官中转移过程中的时空肿瘤-宿主相互作用。Microsc. Res. Tech. 2011. (c)2011 Wiley Periodicals，Inc.
A major challenge of cancer biology is to visualize the dynamics of the metastatic process in secondary organs at high optical resolution in vivo real-time. Here, we presented intravital, dual-colored imaging of liver metastasis formation from a single cancer cell to metastatic colonies in the living liver of living mice using two photon laser scanning microscopy (TPLSM). Red fluorescent protein expressing murine (SL4) or human (HT29) colorectal cancer cell lines were inoculated to the spleen of green fluorescent protein expressing mice. Intravital TPLSM was performed by exteriorizing and fixing the liver lobe of living mice. This was repeated several times for the long-term imaging of the same mouse. Viable cancer cells in the living liver of living mice were visualized intravitally at a magnification of over 600x. Single cancer cells were arrested within hepatic sinusoids 2 h after injection. Platelet aggregation surrounding a cancer cell was observed, indicating a phenomenon of tumor-cell induced platelet aggregation. Cancer cells were extravasated from hepatic sinusoids to the space of Disse. Protrusions of Kupffer cells surrounding a cancer cell were observed, indicating that Kupffer cells appear to phagocytose cancer cells. SL4 cells formed liver metastatic colonies with extensive stromal reaction. Liver metastases by HT29 cells were observed as a cluster of micrometastatic nodules. High-resolution, dual-colored, real-time visualization of cancer metastasis using intravital TLPSM can help to understand spatiotemporal tumor-host interactions during metastatic processes in the living organs of living animals. Microsc. Res. Tech. 2011. (c) 2011 Wiley Periodicals, Inc.