Abstract:
Super-resolution microscopy (SRM) has transformed our understanding of proteins\' subcellular organization and revealed cellular details down to nanometers, far beyond conventional microscopy. While localization precision is independent of the number of fluorophores attached to a biomolecule, labeling density is a decisive factor for resolving complex biological structures. The average distance between adjacent fluorophores should be less than half the desired spatial resolution for optimal clarity. While this was not a major limitation in recent decades, the success of modern microscopy approaching molecular resolution down to the single-digit nanometer range will depend heavily on advancements in fluorescence labeling. This review highlights recent advances and challenges in labeling strategies for SRM, focusing on site-specific labeling technologies. These advancements are crucial for improving SRM precision and expanding our understanding of molecular interactions.
摘要:
超分辨率显微镜(SRM)改变了我们对蛋白质亚细胞组织的理解,并揭示了细胞细节,直至纳米。远远超出传统的显微镜。虽然定位精度与附着在生物分子上的荧光团的数量无关,标记密度是解析复杂生物结构的决定性因素。为了最佳清晰度,相邻荧光团之间的平均距离应小于所需空间分辨率的一半。虽然这不是最近几十年的主要限制,现代显微镜接近分子分辨率下降到一位数纳米范围的成功将在很大程度上取决于荧光标记的进步。这篇综述重点介绍了SRM标签策略的最新进展和挑战,专注于特定地点的标签技术。这些进步对于提高SRM精度和扩大我们对分子相互作用的理解至关重要。