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<p>Fluorescent nanoparticles offer superior brightness and photostability compared with conventional dyes and proteins. However, their relatively large size and complex surface chemistry limit their utility for imaging nanoscale biostructures and tracking individual proteins in living cells. Here we develop single-chain ultrasmall fluorescent polymer dots (<i>su</i>Pdots) with size below 5 nm, comparable to fluorescent proteins. Fabricated via vitrification of conjugated polymer solutions, <i>su</i>Pdots enable tunable fluorescence as well as high-density, specific labelling of multiple subcellular organelles. We demonstrate nanoscopic imaging of continuous ring structures in clathrin-coated pits as well as multi-target stimulated emission depletion imaging. Thanks to their high brightness, <i>su</i>Pdots enable tracking the individual steps of the kinesin-1 motor protein in living cells using standard spinning-disk fluorescence microscopy, with a 16-nm step size and 50-Hz temporal resolution. These demonstrations establish <i>su</i>Pdots as powerful, versatile fluorescent probes for nanoscale-resolution biomolecular imaging with increased accessibility and efficiency for diverse bio-applications.</p>

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Single-chain ultrasmall fluorescent polymer dots enable nanometre-resolution cellular imaging and single protein tracking

  • 期刊论文

  • 发表日期：

    2025年10月3日

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  • Hongwei Yang,
  • Zequan Yan,
  • Xiaolong Liu,
  • Weifeng Liu,
  • Panru Lin,
  • Han Xue,
  • Yayun Wu,
  • Yifei Jiang,
  • Qingrui Fan,
  • Jianjun Wang,
  • Xiaohong Fang

摘要

Fluorescent nanoparticles offer superior brightness and photostability compared with conventional dyes and proteins. However, their relatively large size and complex surface chemistry limit their utility for imaging nanoscale biostructures and tracking individual proteins in living cells. Here we develop single-chain ultrasmall fluorescent polymer dots (suPdots) with size below 5 nm, comparable to fluorescent proteins. Fabricated via vitrification of conjugated polymer solutions, suPdots enable tunable fluorescence as well as high-density, specific labelling of multiple subcellular organelles. We demonstrate nanoscopic imaging of continuous ring structures in clathrin-coated pits as well as multi-target stimulated emission depletion imaging. Thanks to their high brightness, suPdots enable tracking the individual steps of the kinesin-1 motor protein in living cells using standard spinning-disk fluorescence microscopy, with a 16-nm step size and 50-Hz temporal resolution. These demonstrations establish suPdots as powerful, versatile fluorescent probes for nanoscale-resolution biomolecular imaging with increased accessibility and efficiency for diverse bio-applications.