Murooka Thomas T, Mempel Thorsten R
Center for Immunology and Inflammatory Diseases and Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA.
Methods Mol Biol. 2012;757:247-57. doi: 10.1007/978-1-61779-166-6_16.
Intravital microscopy (IVM) allows for the direct in vivo visualization of dynamic biological processes in their physiological context at high spatial and temporal resolution. Novel nonlinear optical imaging modalities, most prominently multiphoton microscopy, have extended the spectrum of cellular functions amenable to IVM investigation to include migration and cell-cell interactions occurring deep inside the highly light-scattering environments of solid tissues, which had so far been inaccessible to conventional microscopy techniques. This has led to important new insights into immune cell behavior at steady state, as well as their change in behavior during an immune response. Here, we describe in detail a technique that allows for the monitoring of lymphocyte motility in the lymph nodes of mice at the single cell level using multiphoton intravital microscopy (MP-IVM).
活体显微镜检查（IVM）能够在生理环境中以高空间和时间分辨率对动态生物过程进行直接的体内可视化观察。新型非线性光学成像方式，尤其是多光子显微镜，已将适合IVM研究的细胞功能范围扩展到包括在实体组织高度光散射环境深处发生的迁移和细胞间相互作用，而这些是传统显微镜技术迄今为止无法触及的。这为稳态下免疫细胞行为以及免疫反应期间其行为变化带来了重要的新见解。在此，我们详细描述一种使用多光子活体显微镜（MP-IVM）在单细胞水平监测小鼠淋巴结中淋巴细胞运动性的技术。
