Renaissance of morphological studies: the examination of functional structures in living animal organs using the in vivo cryotechnique.
Renaissance of morphological studies: the examination of functional structures in living animal organs using the in vivo cryotechnique.
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形态学研究的复兴：使用体内冷冻技术检查活体动物器官的功能结构。
DOI：
10.1007/s12565-016-0355-9
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发表时间：
2016
影响因子：
1.2
通讯作者：
Terada N
中科院分区：
文献类型：
--
作者：
Ohno S;Saitoh Y;Ohno N;Terada N
医学和生物科学家希望了解构成活体动物器官的细胞和组织的体内结构，以及它们的分子成分的位置。最近，通过基因操作产生的荧光标记蛋白质对动物细胞和组织的活体成像已经变得越来越普遍。因此，重要的是要确保从活体动物器官的组织学或免疫组织化学组织切片获得的结果与从相同器官的实时图像获得的结果相一致，这些结果可以使用最近开发的数字成像技术进行评估。在过去的二十年里，我们已经进行了免疫组织化学和形态学研究的细胞和组织在活的动物器官使用一种新的体内冷冻技术。在这种冷冻技术中使用一种特别设计的带或不带冷冻刀的液体冷冻剂系统，解决了在光或电子显微镜检查之前采用的常规制备方法中不可避免地出现的技术问题。我们的体内冷冻技术已被发现是非常有用的逮捕短暂的生理过程中的细胞和组织，并保持其功能成分，如快速变化的信号分子，膜通道，或受体在原位。本文综述了冷冻技术的基本原理和我们在体冷冻技术的意义。此外，它还描述了各种形态学或免疫组织化学发现，使用量子点进行的观察，以及用于评估流经肠组织的红细胞中氧饱和度的基于拉曼低温显微镜的方法。
Medical and biological scientists wish to understand the in vivo structures of the cells and tissues that make up living animal organs, as well as the locations of their molecular components. Recently, the live imaging of animal cells and tissues with fluorescence-labeled proteins produced via gene manipulation has become increasingly common. Therefore, it is important to ensure that findings derived from histological or immunohistochemical tissue sections of living animal organs are compatible with those obtained from live images of the same organs, which can be assessed using recently developed digital imaging techniques. Over the past two decades, we have performed immunohistochemical and morphological studies of the cells and tissues in living animal organs using a novel in vivo cryotechnique. The use of a specially designed liquid cryogen system with or without a cryoknife during this cryotechnique solved the technical problems that inevitably arise during the conventional preparation methods employed prior to light or electron microscopic examinations. Our in vivo cryotechnique has been found to be extremely useful for arresting transient physiological processes in cells and tissues and for maintaining their functional components—such as rapidly changing signaling molecules, membrane channels, or receptors—in situ. The purpose of the present review is to describe the basic mechanism underlying cryotechniques and the significance of our in vivo cryotechnique. In addition, it describes various morphological or immunohistochemical findings, observations made using quantum dots, and a Raman cryomicroscopy-based method for assessing oxygen saturation in the erythrocytes flowing through intestinal tissues.