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Highly Multiplexed, Super-resolution Imaging of T Cells Using madSTORM
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Nanometer-scale Multiplexed Super-Resolution Imaging with an Economic 3D-DNA-PAINT Microscope.

Alexander Auer1,2, Thomas Schlichthaerle1,2, Johannes B Woehrstein1,2

  • 1Faculty of Physics and Center for Nanoscience, LMU Munich, Geschwister-Scholl-Platz 1, 80539, Munich.

Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry
|September 13, 2018
PubMed
Summary

We developed an affordable, custom 3D super-resolution microscope combined with DNA-PAINT imaging. This system enables high-resolution biological imaging, overcoming cost and complexity barriers for broader laboratory adoption.

Keywords:
DNAnanotechnologyphotophysicssingle-molecule microscopysuper-resolution imaging

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Area of Science:

  • Biomedical Imaging
  • Optical Microscopy
  • Nanotechnology

Background:

  • Optical super-resolution microscopy offers molecular contrast and sub-diffraction resolution for biological studies.
  • Widespread adoption is limited by complex sample preparation, image acquisition, and high instrumentation costs.

Purpose of the Study:

  • Introduce a cost-effective, custom-built 3D single-molecule microscope (liteTIRF).
  • Combine liteTIRF with DNA-PAINT (DNA points accumulation for imaging in nanoscale topography) for super-resolution imaging.
  • Demonstrate the system's performance and applicability for biological samples.

Main Methods:

  • Developed a custom-built, economic 3D single-molecule microscope (liteTIRF).
  • Integrated the DNA-PAINT super-resolution technique with the liteTIRF microscope.
  • Validated the system using synthetic DNA origami structures and cellular imaging.

Main Results:

  • The liteTIRF microscope is significantly more affordable than commercial systems.
  • The combined system achieves high-resolution imaging of nanoscale structures.
  • Demonstrated successful single- and multiplexed cellular imaging with the developed setup.

Conclusions:

  • The custom liteTIRF microscope combined with DNA-PAINT provides an accessible super-resolution imaging solution.
  • This approach addresses key limitations hindering the routine use of super-resolution microscopy in biological labs.
  • The system shows promise for detailed structural and cellular imaging at the nanoscale.