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GMars-T Enabling Multimodal Subdiffraction Structural and Functional Fluorescence Imaging in Live Cells.

Sheng Wang1, Xuanze Chen1,2,3, Lei Chang1

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

Analytical Chemistry
|May 4, 2018
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Summary
This summary is machine-generated.

Researchers developed GMars-T, a novel green fluorescent protein, for advanced live-cell imaging. This versatile probe enables both super-resolution microscopy and sensitive detection of protein interactions, enhancing biological research.

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Area of Science:

  • Biophysics
  • Cell Biology
  • Microscopy

Background:

  • Multimodal imaging offers enhanced biological insights and cross-validation.
  • Genetically encoded reversibly photoswitchable fluorescent proteins (RSFPs) are crucial for live-cell nanoscopy techniques like RESOLFT and SOFI.
  • Existing RSFPs often have limitations, requiring specific optical properties for different nanoscopy methods.

Purpose of the Study:

  • To develop a novel monomeric bright green RSFP, GMars-T, with versatile imaging capabilities.
  • To demonstrate GMars-T's suitability for both RESOLFT and photochromic SOFI (pcSOFI) live-cell nanoscopy.
  • To create a GMars-T-based biosensor for sensitive and specific detection of protein-protein interactions (PPIs) in live cells.

Main Methods:

  • Development and characterization of the GMars-T RSFP.
  • Application of GMars-T in RESOLFT and pcSOFI super-resolution microscopy.
  • Construction of a bimolecular fluorescence complementation (BiFC) biosensor utilizing GMars-T for PPI detection.

Main Results:

  • GMars-T was successfully developed as a bright, monomeric green RSFP.
  • GMars-T demonstrated efficacy in both RESOLFT and pcSOFI live-cell imaging.
  • The derived GMars-T BiFC biosensor exhibited high specificity and sensitivity for visualizing PPIs in various subcellular locations at 37 °C.

Conclusions:

  • GMars-T is a versatile fluorescent probe suitable for multimodal super-resolution structural imaging.
  • The GMars-T-based biosensor provides a powerful tool for functional imaging of PPIs in live mammalian cells under physiological conditions.