Imaging In Mice With Fluorescent Proteins: From Macro To Subcellular.
Imaging In Mice With Fluorescent Proteins: From Macro To Subcellular.
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DOI：
10.3390/s8021157
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发表时间：
2008-02-22
期刊：
影响因子：
--
通讯作者：
Hoffman RM
中科院分区：
文献类型：
--
作者：
Hoffman RM
荧光蛋白全身成像已被证明是一种强大的技术，在小动物中有许多应用。更亮的红移蛋白质可以使全身成像更加灵敏，因为组织吸收减少，散射减少。例如，一种名为Katushka的新蛋白质已经被分离出来，它是已知最亮的蛋白质，发射波长超过620 nm。这种新的蛋白质提供了非侵入性全身宏观成像的潜力，如肿瘤生长。对于亚细胞成像，为了观察活小鼠中的细胞质和细胞核动力学，癌细胞在细胞核中用绿色荧光蛋白标记，在细胞质中用红色荧光蛋白标记。在腹部皮瓣中，当癌细胞通过各种方式运输或粘附于血管表面时，对它们在血管中的核和胞质行为进行真实的成像。在外渗过程中，实时双色成像显示癌细胞的胞质突起首先离开血管，细胞核沿着胞质突起。细胞质和细胞核在外渗过程中都发生了变形。还对淋巴管中的癌细胞运输进行成像。为了对肿瘤微环境中的癌细胞/基质细胞相互作用以及活体动物细胞水平上的药物反应进行真实的非侵入性成像，我们开发了一种新的可成像三色动物模型。该模型由移植了双色癌细胞的GFP表达小鼠组成。利用双色癌细胞和高灵敏度的小动物成像系统，现在可以在活体小鼠中真实的观察亚细胞动力学。因此，荧光蛋白能够实现宏观和微观成像技术，从而为体内细胞生物学的新领域提供基础。
Whole-body imaging with fluorescent proteins has been shown to be a powerful technology with many applications in small animals. Brighter, red-shifted proteins can make whole-body imaging even more sensitive due to reduced absorption by tissues and less scatter. For example, a new protein called Katushka has been isolated that is the brightest known protein with emission at wavelengths longer than 620 nm. This new protein offers potential for noninvasive whole-body macro imaging such as of tumor growth. For subcellular imaging, to observe cytoplasmic and nuclear dynamics in the living mouse, cancer cells were labeled in the nucleus with green fluorescent protein and with red fluorescent protein in the cytoplasm. The nuclear and cytoplasmic behavior of cancer cells in real time in blood vessels was imaged as they trafficked 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. Cancer cells trafficking in lymphatic vessels was also imaged. To noninvasively image cancer cell/stromal cell interaction in the tumor microenvironment as well as drug response at the cellular level in live animals in real time, we developed a new imageable three-color animal model. The model consists of GFP-expressing mice transplanted with the dual-color cancer cells. With the dual-color cancer cells and a highly sensitive small animal imaging system, subcellular dynamics can now be observed in live mice in real time. Fluorescent proteins thus enable both macro and micro imaging technology and thereby provide the basis for the new field of in vivo cell biology.
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影响因子：
64.5
作者：
Sakaue-Sawano, Asako;Kurokawa, Hiroshi;Miyawaki, Atsushi
通讯作者：
Miyawaki, Atsushi
影响因子：
45.3
作者：
Chen, Helen X.;Mooney, Margaret;Kaplan, Richard S.
通讯作者：
Kaplan, Richard S.
影响因子：
46.9
作者：
Ntziachristos, V;Ripoll, J;Weissleder, R
通讯作者：
Weissleder, R
影响因子：
11.2
作者：
Tsuji, K;Yamauchi, K;Hoffman, RM
通讯作者：
Hoffman, RM
影响因子：
14.8
作者：
Hoffman, Robert M.;Yang, Meng
通讯作者：
Yang, Meng