Meijer Eelco F J, Jeong Han-Sin, Pereira Ethel R, Ruggieri Thomas A, Blatter Cedric, Vakoc Benjamin J, Padera Timothy P
Edwin L. Steele Laboratories, Department of Radiation Oncology, and Cancer Center, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA.
Department of Otorhinolaryngology-Head and Neck Surgery, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, South Korea.
Nat Protoc. 2017 Aug;12(8):1513-1520. doi: 10.1038/nprot.2017.045. Epub 2017 Jul 6.
Chronic imaging windows in mice have been developed to allow intravital microscopy of many different organs and have proven to be of paramount importance in advancing our knowledge of normal and disease processes. A model system that allows long-term intravital imaging of lymph nodes would facilitate the study of cell behavior in lymph nodes during the generation of immune responses in a variety of disease settings and during the formation of metastatic lesions in cancer-bearing mice. We describe a chronic lymph node window (CLNW) surgical preparation that allows intravital imaging of the inguinal lymph node in mice. The CLNW is custom-made from titanium and incorporates a standard coverslip. It allows stable longitudinal imaging without the need for serial surgeries while preserving lymph node blood and lymph flow. We also describe how to build and use an imaging stage specifically designed for the CLNW to prevent (large) rotational changes as well as respiratory movement during imaging. The entire procedure takes approximately half an hour per mouse, and subsequently allows for longitudinal intravital imaging of the murine lymph node and surrounding structures for up to 14 d. Small-animal surgery experience is required to successfully carry out the protocol.
小鼠慢性成像窗口已被开发出来，以实现对许多不同器官的活体显微镜检查，并已被证明在推进我们对正常和疾病过程的认识方面至关重要。一个能够对淋巴结进行长期活体成像的模型系统，将有助于研究在各种疾病环境下免疫反应产生过程中以及荷瘤小鼠转移瘤形成过程中淋巴结内的细胞行为。我们描述了一种慢性淋巴结窗口（CLNW）手术制备方法，该方法可对小鼠腹股沟淋巴结进行活体成像。CLNW是由钛定制而成，并包含一个标准盖玻片。它允许进行稳定的纵向成像，无需连续手术，同时保留淋巴结的血液和淋巴流动。我们还描述了如何构建和使用专门为CLNW设计的成像平台，以防止成像过程中出现（大幅度）旋转变化以及呼吸运动。整个过程每只小鼠大约需要半小时，随后可对小鼠淋巴结及周围结构进行长达14天的纵向活体成像。成功实施该方案需要有小动物手术经验。
