Intravital imaging of gastrointestinal diseases in preclinical models using two-photon laser scanning microscopy.
Intravital imaging of gastrointestinal diseases in preclinical models using two-photon laser scanning microscopy.
复制标题
使用双光子激光扫描显微镜在临床前模型中对胃肠道疾病进行活体成像。
DOI：
10.1007/s00595-012-0283-9
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Kusunoki M.
中科院分区：
文献类型：
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作者：
Tanaka K;Toiyama Y;Inoue Y;Uchida K;Araki T;Mohri Y;Mizoguchi A;Kusunoki M.
双光子激光扫描显微镜(TPLSM)依靠局限于焦平面的双光子激发，由于其能够在较长时间内对生物材料产生高分辨率、更高穿透性的成像而不会出现明显的光漂白，因此已成为生物医学研究的黄金标准。使用TPLSM对腹部器官进行活体(活体)成像长期以来一直是一个技术挑战，因为很难获得具有可接受的心脏和呼吸运动伪影的高质量和更高放大倍数的成像。利用器官稳定系统建立了活体TPLSM方法，用于对绿色荧光蛋白转基因小鼠的腹内器官进行成像。这种方法被进一步改进，用于使用活体TPLSM对同一小鼠模型中的腹部器官进行时间序列成像。这些方法不仅可以在更高的穿透深度观察单个细胞或组织的微环境更长时间，而且还可以在临床前模型中在多个时间点观察同一小鼠的同一器官。本报告介绍了用于生物医学研究的TPLSM的一般原理和特性。此外，还讨论了时间序列活体TPLSM成像在临床前胃肠道疾病中的方法和应用。
Two-photon laser scanning microscopy (TPLSM), relying on two-photon excitation restricted to the focal plane, has become the gold standard in biomedical research because of its ability to produce high-resolution, higher penetrating imaging of biological materials over an extended duration without an significant photobleaching. Intravital (in vivo) imaging using TPLSM for intra-abdominal organs has long been a technical challenge because of the difficulty of achieving high-quality and higher magnification imaging with acceptable cardiac and respiratory motion artifacts. A method of intravital TPLSM was developed using an organ-stabilizing system for imaging the intra-abdominal organs of green fluorescent protein transgenic mice. This method was further refined for the time-series imaging of intra-abdominal organs in the same mouse model using intravital TPLSM. These procedures allow the observation of not only a single cell or tissue microenvironment at a higher penetrating depth for a longer period of time but also to observe the same organ of the same mouse at multiple time points in preclinical models. This report presents the general principles and properties of TPLSM for biomedical research. In addition, the methods and the usefulness of time-series intravital TPLSM imaging for preclinical gastrointestinal diseases is discussed.