Long-term monitoring of intravital biological processes using fluorescent protein-assisted NIR-II imaging.
Long-term monitoring of intravital biological processes using fluorescent protein-assisted NIR-II imaging.
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DOI：
10.1038/s41467-022-34274-w
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发表时间：
2022-11-04
影响因子：
16.6
通讯作者：
Xu, Zhengping
中科院分区：
文献类型：
--
作者：
Chen, Muxiong;Feng, Zhe;Fan, Xiaoxiao;Sun, Jun;Geng, Weihang;Wu, Tianxiang;Sheng, Jinghao;Qian, Jun;Xu, Zhengping
高空间分辨率、低背景和深组织穿透使得近红外II（NIR-II）荧光成像成为活体观察和测量的最关键工具之一。然而，合成的NIR-II荧光团相对较短的保留时间和潜在的毒性限制了它们的长期应用。在这里，我们报告使用红外荧光蛋白（iRFPs）作为体外和体内NIR-II探针，允许延长连续成像（长达15个月）。作为代表性实例，将iRFP 713敲入小鼠基因组以产生转基因模型，从而允许探针的时间和/或空间表达控制。为了证明其在真正诊断背景下的可行性，我们采用了两种肝再生模型，并成功跟踪了一周的过程。性能和监测效果与μCT相当，上级吲哚菁绿色染料。我们还能够有效地观察胰腺，尽管它位于深部，在生理和病理条件下。这些结果表明，iRFP-assisted NIR-II荧光系统适用于监测各种组织和体内生物过程，提供了一个强大的无创长期成像平台。合成NIR-II荧光团的保留时间和毒性限制了其长期应用。在本文中，作者使用红外荧光蛋白（iRFPs）作为NIR-II探针，在小鼠中进行长时间连续成像，并生成转基因小鼠模型，用于肝脏和胰腺成像。
High spatial resolution, low background, and deep tissue penetration have made near-infrared II (NIR-II) fluorescence imaging one of the most critical tools for in vivo observation and measurement. However, the relatively short retention time and potential toxicity of synthetic NIR-II fluorophores limit their long-term application. Here, we report the use of infrared fluorescent proteins (iRFPs) as in vitro and in vivo NIR-II probes permitting prolonged continuous imaging (up to 15 months). As a representative example, iRFP713 is knocked into the mouse genome to generate a transgenic model to allow temporal and/or spatial expression control of the probe. To demonstrate its feasibility in a genuine diagnostic context, we adopt two liver regeneration models and successfully track the process for a week. The performance and monitoring efficacy are comparable to those of μCT and superior to those of indocyanine green dye. We are also able to effectively observe the pancreas, despite its deep location, under both physiological and pathological conditions. These results indicate that the iRFP-assisted NIR-II fluorescence system is suitable for monitoring various tissues and in vivo biological processes, providing a powerful noninvasive long-term imaging platform. The retention time and toxicity of synthetic NIR-II fluorophores limit long-term application. Here the authors use infrared fluorescent proteins (iRFPs) as NIR-II probes for prolonged continuous imaging in mice, and generate a transgenic mouse model which they use to image liver and pancreas.
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作者：
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