Scholarlink

<p>Latest refinements in super-resolution microscopy achieved spatial resolutions in the one nanometer range. However, improvement of localization precision advanced faster than labeling methods impeding the translation of such high resolutions to cells. Hence, imaging of the nano-architecture of endogenous multiprotein complexes remains challenging. Here we introduce an expansion microscopy (ExM) method using double-homogenized hydrogels that enables <i>direct</i> stochastic optical reconstruction microscopy (<i>d</i>STORM) of 7-8-fold expanded immunolabeled samples. The ~4-fold increase in labeling density resolves the 8 nm spacing between neighboring α-tubulin molecules in microtubules, the polyhedral lattice of clathrin-coated pits, and provides evidence for the 8 nm periodicity of α/β-tubulin heterodimers. Two-color Ex-<i>d</i>STORM further reveals the molecular organization of RIM and the synaptic vesicle priming protein Munc13-1 in 44–48 nm ring-like presynaptic structures in neurons. Ex-<i>d</i>STORM thus enables nanometer-resolution imaging of endogenous multiprotein complexes in genetically unmodified cells, providing a versatile tool for studying molecular organization in physiological contexts.</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

Resolving endogenous protein organization in cells with nanometer resolution

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年7月29日

查看全文
  • Janna Eilts,
  • Marvin Jungblut,
  • Dominic A. Helmerich,
  • Stefan Sachs,
  • Christian Werner,
  • Cristian-Alexandru Bogaciu,
  • Ali H. Shaib,
  • Silvio O. Rizzoli,
  • Philip Kollmannsberger,
  • Sören Doose,
  • Markus Sauer

摘要

Latest refinements in super-resolution microscopy achieved spatial resolutions in the one nanometer range. However, improvement of localization precision advanced faster than labeling methods impeding the translation of such high resolutions to cells. Hence, imaging of the nano-architecture of endogenous multiprotein complexes remains challenging. Here we introduce an expansion microscopy (ExM) method using double-homogenized hydrogels that enables direct stochastic optical reconstruction microscopy (dSTORM) of 7-8-fold expanded immunolabeled samples. The ~4-fold increase in labeling density resolves the 8 nm spacing between neighboring α-tubulin molecules in microtubules, the polyhedral lattice of clathrin-coated pits, and provides evidence for the 8 nm periodicity of α/β-tubulin heterodimers. Two-color Ex-dSTORM further reveals the molecular organization of RIM and the synaptic vesicle priming protein Munc13-1 in 44–48 nm ring-like presynaptic structures in neurons. Ex-dSTORM thus enables nanometer-resolution imaging of endogenous multiprotein complexes in genetically unmodified cells, providing a versatile tool for studying molecular organization in physiological contexts.