Schodt David J, Wester Michael J, Fazel Mohamadreza, Khan Sajjad, Mazloom-Farsibaf Hanieh, Pallikkuth Sandeep, Meddens Marjolein B M, Farzam Farzin, Burns Eric A, Kanagy William K, Rinaldi Derek A, Jhamba Elton, Liu Sheng, Relich Peter K, Olah Mark J, Steinberg Stanly L, Lidke Keith A
Department of Physics and Astronomy, University of New Mexico, Albuquerque, United States of America.
Department of Mathematics and Statistics, Albuquerque, University of New Mexico, United States of America.
J Open Source Softw. 2023;8(90). doi: 10.21105/joss.05563. Epub 2023 Oct 2.
Fluorescence single molecule imaging comprises a variety of techniques that involve detecting individual fluorescent molecules. Many of these techniques involve localizing individual fluorescent molecules with precisions below the diffraction limit, which limits the spatial resolution of (visible) light-based microscopes. These methodologies are widely used to image biological structures at the nanometer scale by fluorescently tagging the structures of interest, elucidating details of the biological behavior observed. Two common techniques are single-molecule localization microscopy (SMLM), (Betzig et al., 2006; Fazel & Wester, 2022; Hell, 2007; Lidke et al., 2005; Rust et al., 2006; van de Linde et al., 2011) which is used to produce 2D or 3D super-resolution images of static or nearly static structures, and single-particle tracking (SPT) (Shen et al., 2017), which follows the time course of one or a very small number of moving tagged molecules. SMLM often involves distributions of particles at medium to high density, while SPT works in a very low density domain. These procedures all require intensive numerical computation, and the methods are tightly interwoven.
荧光单分子成像包含多种涉及检测单个荧光分子的技术。这些技术中的许多都涉及以低于衍射极限的精度定位单个荧光分子，而衍射极限限制了基于（可见）光的显微镜的空间分辨率。通过对感兴趣的结构进行荧光标记，这些方法被广泛用于对纳米尺度的生物结构进行成像，从而阐明所观察到的生物行为的细节。两种常见的技术是单分子定位显微镜（SMLM）（贝齐格等人，2006年；法泽尔和韦斯特，2022年；赫尔，2007年；利德克等人，2005年；拉斯特等人，2006年；范德林德等人，2011年），用于生成静态或近乎静态结构的二维或三维超分辨率图像，以及单粒子追踪（SPT）（沈等人，2017年），它跟踪一个或极少数移动标记分子的时间进程。SMLM通常涉及中到高密度的粒子分布，而SPT在非常低密度的领域中起作用。这些程序都需要密集的数值计算，并且这些方法紧密交织在一起。
