Expansion Microscopy with Multifunctional Polymer Dots

Jie Liu, Xiaofeng Fang, Zhihe Liu, Rongqin Li, Yicheng Yang, Yujie Sun, Zhongying Zhao, Changfeng Wu*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Expansion microscopy (ExM) provides nanoscale resolution on conventional microscopes via physically enlarging specimens with swellable polyelectrolyte gels. However, challenges involving fluorophore degradation and dilution during sample expansion have yet to be overcome. Herein, sequential cellular targeting, gel anchoring, and high-fidelity fluorescence reported using multifunctional polymer dots (Pdots) designed for ExM applications are demonstrated. The impressive brightness of the Pdots facilitates multicolor ExM, thereby enabling visualization of a variety of subcellular structures and neuron synapses. The average fluorescence intensities of Pdots in ExM range from ≈3 to 6 times higher than those achieved using commercially available Alexa dyes. Moreover, the fluorescence brightness and optical fluctuation are significantly improved by a surfactant-containing expansion buffer, which enables further resolution enhancement via super-resolution optical fluctuation imaging (SOFI). The combination of ExM and SOFI allows subcellular structures of ≈30 nm to be resolved by conventional microscopes. These results highlight the immense potential of multifunctional Pdots for ExM-enhanced super-resolution imaging.
Original languageEnglish
Article number2007854
JournalAdvanced Materials
Volume33
Issue number25
Early online date14 May 2021
DOIs
Publication statusPublished - 24 Jun 2021

Scopus Subject Areas

  • Materials Science(all)
  • Mechanics of Materials
  • Mechanical Engineering

User-Defined Keywords

  • expansion microscopy
  • fluorescent probes
  • polymer dots
  • subcellular labeling
  • super-resolution optical fluctuation microscopy

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