Yan Guangjie, Zhang Yuteng, Allamprese Alexis, Brooks Kameron N, Chen Wenkai, Yan Shudan, Chen Tai-Yen
Department of Chemistry, University of Houston, Houston, Texas 77204.
J Chem Educ. 2024 Feb 13;101(2):514-520. doi: 10.1021/acs.jchemed.3c00938. Epub 2024 Jan 5.
Single-molecule localization microscopy (SMLM) has revolutionized our ability to visualize cellular structures, offering unprecedented detail. However, the intricate biophysical principles that underlie SMLM can be daunting for newcomers, particularly undergraduate and graduate students. To address this challenge, we introduce the fundamental concepts of SMLM, providing a solid theoretical foundation. In addition, we have developed an intuitive graphical interface APP that simplifies these core concepts, making them more accessible for students. This APP clarifies how super-resolved images are fitted and highlights the crucial factors determining image quality. Our approach deepens students' understanding of SMLM by combining theoretical instruction with practical learning. This development equips them with the skills to carry out single-molecule super-resolved experiments and explore the microscopic world beyond the diffraction limit.
单分子定位显微镜(SMLM)彻底改变了我们可视化细胞结构的能力,提供了前所未有的细节。然而,SMLM背后复杂的生物物理原理对于新手,尤其是本科生和研究生来说可能令人生畏。为应对这一挑战,我们介绍了SMLM的基本概念,提供了坚实的理论基础。此外,我们开发了一个直观的图形界面应用程序,简化了这些核心概念,使其对学生来说更容易理解。这个应用程序阐明了超分辨图像是如何拟合的,并突出了决定图像质量的关键因素。我们的方法通过将理论指导与实践学习相结合,加深了学生对SMLM的理解。这一进展使他们具备了进行单分子超分辨实验的技能,并能够探索超越衍射极限的微观世界。