Development of real-time subcellular dynamic multicolor imaging of cancer-cell trafficking in live mice with a variable-magnification whole-mouse imaging system
Development of real-time subcellular dynamic multicolor imaging of cancer-cell trafficking in live mice with a variable-magnification whole-mouse imaging system
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DOI：
10.1158/0008-5472.can-05-3927
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发表时间：
2006-04-15
期刊：
影响因子：
11.2
通讯作者：
Hoffman, RM
中科院分区：
文献类型：
--
作者：
Yamauchi, K;Yang, M;Hoffman, RM
利用双色荧光细胞和高灵敏度的宏观光学和微观光学全鼠成像系统，我们在这里报道了活体小鼠癌细胞运输亚细胞实时成像的发展。用绿色荧光蛋白在细胞核内标记肿瘤细胞，用红色荧光蛋白在细胞质内标记肿瘤细胞，观察活体小鼠细胞质和细胞核的动态变化。通过血管途径在裸鼠腹部皮瓣中注射双色癌细胞。采用Olympus OV100全鼠成像系统对小鼠进行成像，该成像系统带有灵敏的CCD相机和五个物镜，中心和共焦，从大细胞到亚细胞进行成像。我们实时观察了癌细胞在血管中通过各种方式移动或粘附在腹部皮瓣血管表面时的核和细胞质行为。在外渗过程中，实时双色成像显示癌细胞的细胞质突首先离开血管，细胞核沿着细胞质突起。外渗过程中细胞质和细胞核均发生变形。不同癌细胞系的外渗能力似乎有很大差异。利用双色癌细胞和高灵敏度的全鼠成像系统，现在可以在活体小鼠中实时观察癌症转移的亚细胞动力学。该成像技术将有助于进一步了解转移的关键步骤，并为抗转移药物的开发提供可见靶点。
With the use of dual-color fluorescent cells and a highly sensitive whole-mouse imaging system with both macro-optics and micro-optics, we report here the development of subcellular real-time imaging of cancer cell trafficking in live mice. To observe cytoplasmic and nuclear dynamics in the living mouse, tumor cells were labeled in the nucleus with green fluorescent protein and with red fluorescent protein in the cytoplasm. Dual-color cancer cells were injected by a vascular route in an abdominal skin flap in nude mice. The mice were imaged with an Olympus OV100 whole-mouse imaging system with a sensitive CCD camera and five objective lenses, parcentered and parfocal, enabling imaging from macrocellular to subcellular. We observed the nuclear and cytoplasmic behavior of cancer cells in real time in blood vessels as they moved by various means or adhered to the vessel surface in the abdominal skin flap. During extravasation, real-time dual-color imaging showed that cytoplasmic processes of the cancer cells exited the vessels first, with nuclei following along the cytoplasmic projections. Both cytoplasm and nuclei underwent deformation during extravasation. Different cancer cell lines seemed to strongly vary in their ability to extravasate. With the dual-color cancer cells and the highly sensitive whole-mouse imaging system described here, the subcellular dynamics of cancer metastasis can now be observed in live mice in real time. This imaging technology will enable further understanding of the critical steps of metastasis and provide visible targets for antimetastasis drug development.