Netherlands Cancer Institute, Amsterdam, Netherlands.
Methods Mol Biol. 2021;2350:69-76. doi: 10.1007/978-1-0716-1593-5_5.
Super Resolution (SR) microscopy has become a powerful tool to study cellular architecture at the nanometer scale. Single molecule localization microscopy (SMLM) is a method in which fluorophore labels repeatedly switch On and Off ("blink"). Their exact locations are estimated by computing the centers of individual blinks. Therefore, the image quality depends on the density of the detected labels, as well as the accuracy of the estimation of their location. Both are influenced by several factors. Here we present a step-by-step method that optimizes many of these factors to facilitate multicolor imaging.
超分辨率（SR）显微镜已成为研究纳米级细胞结构的有力工具。单分子定位显微镜（SMLM）是一种荧光团标记物反复开启和关闭（“闪烁”）的方法。通过计算单个闪烁的中心来估计它们的确切位置。因此，图像质量取决于检测到的标记的密度，以及其位置估计的准确性。这两个因素都受到多个因素的影响。本文提供了一种逐步的方法，可以优化这些因素中的许多因素，以促进多色成像。
