Subcellular imaging of cancer cells in live mice.
Subcellular imaging of cancer cells in live mice.
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DOI：
10.1007/978-1-59745-549-7_9
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发表时间：
2007
影响因子：
--
通讯作者：
R. Hoffman
中科院分区：
文献类型：
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作者：
R. Hoffman
双色荧光细胞，一种颜色在细胞核中，另一种颜色在细胞质中，可以在体内和体外活细胞中实现实时核-细胞质动态可视化。为了获得双色细胞，红色荧光蛋白（RFP）在癌细胞的细胞质中表达，而与组蛋白H2B连接的绿色荧光蛋白（GFP）在细胞核中表达。核 GFP 表达可以实现核动力学的可视化，而同步细胞质 RFP 表达可以实现核质比以及同时细胞和核形状变化的可视化。这种方法使我们能够证明毛细血管中的癌细胞和癌细胞核会伸长以适应这些血管的宽度。毛细血管中癌细胞主轴的平均长度增加到正常长度的大约四倍。原子核的长度增加了 1.6 倍。直径超过 8 μm 的毛细血管中的癌细胞迁移速度高达 48.3 μm/h。利用双色荧光细胞和奥林巴斯OV100这种具有宏观光学和微观光学的高灵敏度全小鼠成像系统，可以实现活体小鼠癌细胞运输的亚细胞实时成像。外渗也可以实时成像。双色成像显示癌细胞的细胞质过程首先离开血管，细胞核沿着细胞质的突出部分排出。双色体内细胞成像还用于可视化癌细胞注射到小鼠门静脉后的运输、核细胞质动力学和活力。
Dual-color fluorescent cells, with one color in the nucleus and the other in the cytoplasm, enable real-time nuclear-cytoplasmic dynamics to be visualized in living cells in vivo as well as in vitro. To obtain the dual-color cells, red fluorescent protein (RFP) is expressed in the cytoplasm of cancer cells, and green fluorescent protein (GFP) linked to histone H2B is expressed in the nucleus. Nuclear GFP expression allows visualization of nuclear dynamics, whereas simultaneous cytoplasmic RFP expression allows visualization of nuclear-cytoplasmic ratios as well as simultaneous cell and nuclear shape changes. This methodology has allowed us to show that the cells and nuclei of cancer cells in the capillaries elongate to fit the width of these vessels. The average length of the major axis of the cancer cells in the capillaries increased to approximately four times their normal length. The nuclei increased their length 1.6 times. Cancer cells in capillaries over 8 μm in diameter were shown to migrate at up to 48.3 μm/h. With the use of dual-color fluorescent cells and the Olympus OV100, a highly sensitive whole-mouse imaging system with both macrooptics and microoptics, it is possible to achieve subcellular real-time imaging of cancer cell trafficking in live mice. Extravasation can also be imaged in real time. Dual-color imaging showed that cytoplasmic processes of cancer cells exited the vessels first, with nuclei following along the cytoplasmic projections. Dual-color in vivo cellular imaging was also used to visualize trafficking, nuclear-cytoplasmic dynamics, and the viability of cancer cells after their injection into the portal vein of mice.