Barr Valarie A, Yi Jason, Samelson Lawrence E
Laboratory of Cellular and Molecular Biology, Center for Cancer Research, National Cancer Institute, Bethesda, MD, 20892-4256, USA.
Methods Mol Biol. 2017;1584:183-206. doi: 10.1007/978-1-4939-6881-7_13.
Single-molecule localization microscopy (SMLM) comprises methods that produce super-resolution images from molecular locations of single molecules. These techniques mathematically determine the center of a diffraction-limited spot produced by a fluorescent molecule, which represents the most likely location of the molecule. Only a small cohort of well-separated molecules is visualized in a single image, and then many images are obtained from a single sample. The localizations from all the images are combined to produce a super-resolution picture of the sample. Here we describe the application of two methods, photoactivation localization microscopy (PALM) and direct stochastic optical reconstruction microscopy (dSTORM), to the study of signaling microclusters in T cells.
单分子定位显微镜（SMLM）包含从单分子的分子位置生成超分辨率图像的方法。这些技术通过数学方法确定荧光分子产生的衍射极限光斑的中心，该中心代表分子最可能的位置。在单幅图像中仅能看到一小群分离良好的分子，然后从单个样本中获取多幅图像。将所有图像中的定位信息合并起来，以生成样本的超分辨率图像。在此，我们描述两种方法，即光激活定位显微镜（PALM）和直接随机光学重建显微镜（dSTORM）在T细胞信号微簇研究中的应用。
