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Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
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Related Experiment Video

Updated: Jun 24, 2025

Visualization of the Immunological Synapse by Dual Color Time-gated Stimulated Emission Depletion STED Nanoscopy
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OpenSTED: open-source dynamic intensity minimum system for stimulated emission depletion microscopy.

Stephanie A Pierce1, Jordan Jacobelli2,3, Katherine S Given4

  • 1University of Colorado Anschutz Medical Campus, Department of Bioengineering, Aurora, Colorado, United States.

Neurophotonics
|June 13, 2024
PubMed
Summary
This summary is machine-generated.

We developed an open-source Dynamic Intensity Minimum (DyMIN) system to reduce photobleaching in Stimulated Emission Depletion (STED) microscopy. This cost-effective add-on enables high-resolution, long-term live-cell imaging with improved signal-to-noise ratios.

Keywords:
dynamic minimumfluorescencemicroscopystimulated emission depletionsuper-resolution

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Area of Science:

  • Microscopy and Imaging Technologies
  • Biophysics
  • Cell Biology

Background:

  • Stimulated Emission Depletion (STED) microscopy offers super-resolution imaging but is limited by high laser power, causing photobleaching and sample damage.
  • Existing STED techniques struggle with sensitive biological samples, restricting long-term live imaging and high-resolution data acquisition.
  • A commercially available stand-alone module for Dynamic Intensity Minimum (DyMIN) to mitigate these issues is currently unavailable.

Purpose of the Study:

  • To develop and demonstrate an open-source Dynamic Intensity Minimum (DyMIN) system for reducing photobleaching in STED microscopy.
  • To enable enhanced time-lapse imaging of live biological samples with improved resolution and reduced photodamage.
  • To provide an accessible and cost-effective solution for researchers using STED microscopy.

Main Methods:

  • An open-source, three-step DyMIN system was designed and implemented on a STED microscope.
  • The system utilizes a fast multiplexer circuit and an inexpensive field-programmable gate array controlled by Labview software.
  • Software and circuit diagrams were made freely available as a stand-alone module for STED microscopes.

Main Results:

  • The custom DyMIN system demonstrated significantly reduced photobleaching in time-lapse imaging of beads, cells, and tissue samples.
  • A higher signal was recorded for bead samples using DyMIN compared to conventional STED after a 50-image sequence.
  • The system proved effective for time-lapse STED imaging of live cells and brain tissue slices, showcasing its versatility.

Conclusions:

  • The developed open-source DyMIN system is an inexpensive add-on that effectively reduces photobleaching in conventional STED microscopes.
  • This system significantly improves signal-to-noise ratios, facilitating dynamic time-lapse STED imaging of live samples.
  • The accessibility of the design empowers researchers to enhance their STED imaging capabilities without commercial limitations.