Kim Young Min, Jeong Soi, Choe Young Ho, Hyun Young-Min
Department of Anatomy and Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Korea.
Acute Crit Care. 2019 May;34(2):101-107. doi: 10.4266/acc.2019.00542. Epub 2019 May 31.
Two-photon intravital imaging is a powerful method by which researchers are able to directly observe biological phenomena in live organisms. Researchers in various biomedical research fields have applied two-photon imaging to a variety of target organs by utilizing this technology's ability to penetrate to significant depths with minimal phototoxicity. The mouse respiratory system in inflammation models is a good example, as two-photon intravital imaging can provide insights as to how the immune system is activated in response to inflammation within the respiratory system. Inflammation models can be generated via influenza viral, bacterial, or lipopolysaccharide injection. To exteriorize the lungs or trachea, thoracotomy or tracheotomy is performed, respectively; the appropriate combination of inflammation induction and organ exposure is selected depending on the study purpose. On the other hand, visualizing the movement of leukocytes is also an important component; to this end, immune cell populations of interest are either labeled via the genetic attachment of fluorescent proteins or stained with antibodies or dyes. With the proper selection of methods at each step, twophoton intravital imaging can yield visual evidence regarding immune responses to inflammation.
双光子活体成像技术是一种强大的方法，通过该方法研究人员能够直接观察活生物体中的生物学现象。各个生物医学研究领域的研究人员利用这项技术能够在最小光毒性的情况下穿透到相当深度的能力，将双光子成像应用于各种靶器官。炎症模型中的小鼠呼吸系统就是一个很好的例子，因为双光子活体成像可以提供关于免疫系统如何响应呼吸系统内的炎症而被激活的见解。炎症模型可以通过注射流感病毒、细菌或脂多糖来建立。为了暴露肺部或气管，分别进行开胸手术或气管切开术；根据研究目的选择炎症诱导和器官暴露的适当组合。另一方面，可视化白细胞的运动也是一个重要组成部分；为此，感兴趣的免疫细胞群体要么通过荧光蛋白的基因附着进行标记，要么用抗体或染料进行染色。通过在每个步骤中正确选择方法，双光子活体成像可以产生关于炎症免疫反应的视觉证据。
