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Expansion enhanced nanoscopy.

Rongqin Li1, Xuanze Chen, Zixi Lin

  • 1State Key Laboratory of Membrane Biology, Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences, Peking University, Beijing 100871, China. chenxuanze@pku.edu.cn sun_yujie@pku.edu.cn.

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|September 19, 2018
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Summary
This summary is machine-generated.

New expansion nanoscopic imaging (ExN) methods overcome fluorescence loss in expansion microscopy (ExM). This enables super-resolution techniques like STED and SOFI to achieve higher resolution for cellular and tissue imaging.

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Area of Science:

  • Biophysics
  • Microscopy
  • Cell Biology

Background:

  • Optical super-resolution fluorescence microscopy offers unprecedented views of subcellular structures.
  • Expansion microscopy (ExM) enhances resolution by physically expanding samples.
  • Combining ExM with super-resolution techniques is promising but hindered by signal loss during sample preparation.

Purpose of the Study:

  • To develop strategies for retaining and increasing fluorescence in expanded samples for super-resolution imaging.
  • To enable expansion nanoscopic imaging (ExN) approaches by achieving sufficient labeling density for techniques like STED and SOFI.

Main Methods:

  • Developed novel labeling strategies to enhance fluorescence signal in ExM samples.
  • Applied these strategies to achieve Nyquist sampling criteria for super-resolution imaging.
  • Integrated ExM with stimulated emission depletion (STED) and super-resolution optical fluctuation imaging (SOFI) to create ExSTED and ExSOFI.

Main Results:

  • Successfully retained and increased fluorescence in expanded samples.
  • Achieved labeling density suitable for super-resolution imaging.
  • ExSTED and ExSOFI demonstrated up to a 4-fold resolution enhancement compared to standard STED and SOFI.

Conclusions:

  • The developed ExN approaches (ExSTED, ExSOFI) effectively address fluorescence signal loss in ExM.
  • These methods provide a simple and effective way to achieve high-resolution imaging at cellular and tissue levels.
  • ExN significantly enhances the capabilities of existing super-resolution microscopy techniques.