From Molecules to Classrooms: A Comprehensive Guide to Single-Molecule Localization Microscopy
From Molecules to Classrooms: A Comprehensive Guide to Single-Molecule Localization Microscopy
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DOI：
10.1021/acs.jchemed.3c00938
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发表时间：
2024-01-05
影响因子：
3
通讯作者：
Chen，Tai-Yen
中科院分区：
文献类型：
--
作者：
Yan，Guangjie;Zhang，Yuteng;Chen，Tai-Yen
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可能会让新手望而却步，特别是本科生和研究生。为了应对这一挑战，我们介绍了SMLM的基本概念，为我们提供了坚实的理论基础。此外，我们还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，使学生更容易理解它们。这款应用程序阐明了超分辨率图像是如何匹配的，并突出了决定图像质量的关键因素。我们的方法通过理论教学和实践学习相结合的方式加深了学生对SMLM的理解。这一进展使他们具备了进行单分子超分辨实验和探索超出衍射极限的微观世界的技能。
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 and makes them more accessible for students. This APP clarifies how super-resolved images are fitted and highlights the crucial factors that determine 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.