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Super-resolution Imaging of the Bacterial Division Machinery
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Sequential super-resolution imaging using DNA strand displacement.

Sandeep Pallikkuth1, Cheyenne Martin2,3, Farzin Farzam1

  • 1Department of Physics and Astronomy, University of New Mexico, Albuquerque, New Mexico, United States of America.

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|September 1, 2018
PubMed
Summary
This summary is machine-generated.

This study introduces a rapid DNA strand displacement method for sequential fluorescence labeling and removal, enabling multiplexed super-resolution imaging of cellular structures with a single fluorophore. This technique simplifies super-resolution microscopy and avoids chromatic aberrations.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Microscopy

Background:

  • Sequential labeling and imaging in fluorescence microscopy allows multiplexed visualization of cellular structures using a single fluorophore species.
  • Super-resolution microscopy benefits from simplified optical setups and elimination of chromatic aberrations when using a single fluorophore.
  • Existing methods for sequential imaging can be time-consuming and complex.

Purpose of the Study:

  • To develop a rapid and user-friendly method for sequential fluorophore labeling and removal.
  • To enable multiplexed super-resolution imaging of subcellular structures.
  • To demonstrate the applicability of DNA strand displacement for dynamic labeling in microscopy.

Main Methods:

  • A DNA strand displacement technique was employed for site-specific labeling and fluorophore removal.
  • Antibody-bound DNA tags were hybridized with complementary, dye-labeled DNA strands for structure visualization.
  • Toehold-mediated strand displacement was used to remove the dye-labeled strands after imaging, followed by washing.

Main Results:

  • The developed method allows for quick labeling and removal of fluorophores, with each cycle taking only a few minutes.
  • Successful demonstration of sequential DNA-based labeling and removal for multiplexed imaging.
  • Application of the method in dSTORM (direct Stochastic Optical Reconstruction Microscopy) for super-resolution imaging of subcellular structures.

Conclusions:

  • DNA strand displacement offers an efficient and versatile approach for sequential fluorophore manipulation in microscopy.
  • This method facilitates simplified super-resolution imaging workflows, enabling multiplexed visualization of multiple targets within the same cell.
  • The technique holds promise for advancing high-content imaging and cellular structure analysis.