Scholarlink

<p>Single-molecule localization microscopy (SMLM) has transformed biological imaging by enabling nanoscale visualization of intricate subcellular structures. However, conventional three-dimensional SMLM techniques typically exhibit lower axial resolution than lateral resolution, hindering isotropic investigations. Interferometric approaches, such as 4Pi-SMLM, enhance axial resolution by approximately fivefold through dual-objective coherent fluorescence detection, surpassing lateral resolution. Here we present 4Pi-SIMFLUX, which integrates structured illumination into 4Pi-SMLM to double its lateral resolution, achieving near-isotropic three-dimensional localization precision of 2–3 nm. We demonstrate that 4Pi-SIMFLUX breaks the 10-nm resolution barrier in biological samples, resolving microtubule ultrastructure and nuclear pore complexes with exceptional detail and clarity, while accounting for label size and localization density. Furthermore, it enables simultaneous multicolor imaging for interrogating multiple cellular components and high-fidelity, whole-cell visualization that captures comprehensive spatial organization. 4Pi-SIMFLUX effectively bridges the axial–lateral resolution gap, establishing a robust tool for molecular-scale imaging in complex cellular environments.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

4Pi-SIMFLUX: 4Pi single-molecule localization microscopy with structured illumination

  • 期刊论文

  • 发表日期：

    2025年11月20日

查看全文
  • Qian Wang,
  • Bei Zheng,
  • Zijing Yu,
  • Yajing Zhan,
  • Qiuyang Dai,
  • Xulong Wang,
  • Shuxin Li,
  • Yu Qiao,
  • Linlin Chen,
  • Xiaochun Yu,
  • Yu Lin,
  • Yongdeng Zhang

摘要

Single-molecule localization microscopy (SMLM) has transformed biological imaging by enabling nanoscale visualization of intricate subcellular structures. However, conventional three-dimensional SMLM techniques typically exhibit lower axial resolution than lateral resolution, hindering isotropic investigations. Interferometric approaches, such as 4Pi-SMLM, enhance axial resolution by approximately fivefold through dual-objective coherent fluorescence detection, surpassing lateral resolution. Here we present 4Pi-SIMFLUX, which integrates structured illumination into 4Pi-SMLM to double its lateral resolution, achieving near-isotropic three-dimensional localization precision of 2–3 nm. We demonstrate that 4Pi-SIMFLUX breaks the 10-nm resolution barrier in biological samples, resolving microtubule ultrastructure and nuclear pore complexes with exceptional detail and clarity, while accounting for label size and localization density. Furthermore, it enables simultaneous multicolor imaging for interrogating multiple cellular components and high-fidelity, whole-cell visualization that captures comprehensive spatial organization. 4Pi-SIMFLUX effectively bridges the axial–lateral resolution gap, establishing a robust tool for molecular-scale imaging in complex cellular environments.