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首页> 外文会议>Conference on Small Animal Whole-Body Optical Imaging Based on Genetically Engineered Probes; 20080121-22; San Jose,CA(US) >Subcellular Real-Time In Vivo Imaging of Intralymphatic and Intravascular Cancer-Cell Trafficking
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Subcellular Real-Time In Vivo Imaging of Intralymphatic and Intravascular Cancer-Cell Trafficking

机译：淋巴内和血管内癌细胞转移的亚细胞实时体内成像

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With the use of fluorescent cells labeled with green fluorescent protein (GFP) in the nucleus and red fluorescent protein (RFP) in the cytoplasm and a highly sensitive small animal imaging system with both macro-optics and micro-optics, we have developed subcellular real-time imaging of cancer cell trafficking in live mice. 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 small animal imaging system with a sensitive CCD camera and four 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. We have also developed real-time imaging of cancer cell trafficking in lymphatic vessels. Cancer cells labeled with GFP and/or RFP were injected into the inguinal lymph node of nude mice. The labeled cancer cells trafficked through lymphatic vessels where they were imaged via a skin flap in real-time at the cellular level until they entered the axillary lymph node. The bright dual-color fluorescence of the cancer cells and the real-time microscopic imaging capability of the Olympus OV100 enabled imaging the trafficking cancer cells in both blood vessels and lymphatics. With the dual-color cancer cells and the highly sensitive imaging system described here, the subcellular dynamics of cancer metastasis can now be observed in live mice in real time.
机译：利用在细胞核中标记有绿色荧光蛋白（GFP）和在细胞质中标记有红色荧光蛋白（RFP）的荧光细胞以及具有宏观和微观光学特性的高度敏感的小动物成像系统，我们开发了亚细胞真活细胞中癌细胞运输的实时成像。通过血管途径将双色癌细胞注射到裸鼠的腹部皮瓣中。使用奥林巴斯OV100小动物成像系统对小鼠进行成像，该系统具有灵敏的CCD照相机和四个物镜（同心和近焦点），能够从大细胞到亚细胞成像。我们观察到癌细胞通过各种方式移动或粘附在腹部皮肤皮瓣的血管表面中时，实时在血管中观察其核和细胞质行为。在外渗过程中，实时双色成像显示癌细胞的细胞质过程首先离开血管，随后细胞核沿着细胞质投影。在外渗过程中，细胞质和细胞核均发生变形。不同的癌细胞系似乎在其扩散能力上有很大差异。我们还开发了癌细胞在淋巴管中运输的实时成像。将标记有GFP和/或RFP的癌细胞注入裸鼠的腹股沟淋巴结。标记的癌细胞通过淋巴管运输，并通过皮瓣在细胞水平实时成像，直到进入腋窝淋巴结。癌细胞的明亮双色荧光和Olympus OV100的实时显微成像功能可对血管和淋巴管中的运输癌细胞进行成像。使用此处描述的双色癌细胞和高度敏感的成像系统，现在可以实时在活小鼠中观察到癌症转移的亚细胞动力学。

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