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Super-resolution Fluorescence Microscopy

Super-resolution Fluorescence Microscopy

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: Feb 23, 2026

Visualization of the Immunological Synapse by Dual Color Time-gated Stimulated Emission Depletion STED Nanoscopy

Visualization of the Immunological Synapse by Dual Color Time-gated Stimulated Emission Depletion STED Nanoscopy

Published on: March 24, 2014

Adaptive-illumination STED nanoscopy.

Jörn Heine¹, Matthias Reuss¹, Benjamin Harke¹

¹Abberior Instruments GmbH, 37077 Göttingen, Germany.

Proceedings of the National Academy of Sciences of the United States of America

|August 30, 2017

Summary

This summary is machine-generated.

Dynamic Intensity Minimum (DyMIN) scanning reduces light exposure in super-resolution microscopy. This method dynamically adjusts light intensity, significantly lowering photobleaching and enabling brighter images with minimal sample damage.

Keywords:

STED microscopy adaptive illumination fluorescence nanoscopy photobleaching superresolution

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Area of Science:

Super-resolution microscopy
Biophysics
Optical imaging

Background:

STED/RESOLFT microscopy uses light-driven molecular state transitions for super-resolution imaging.
High light intensities are typically required, leading to increased photobleaching, especially in sparse regions.

Purpose of the Study:

To introduce Dynamic Intensity Minimum (DyMIN) scanning to reduce sample exposure in STED/RESOLFT microscopy.
To generalize and expand upon RESCue and MINFIELD concepts for optimized light dose management.

Main Methods:

DyMIN scanning dynamically adjusts light intensity based on local feature density.
The intensity is tuned per pixel/voxel to match the required resolution and minimize unnecessary light exposure.
Fluorescence is recorded only where fluorophores are detected within a sub-resolution neighborhood.

Main Results:

DyMIN scanning reduces STED light dose by up to ~20-fold in common biological imaging.
For sparser 2D and 3D samples, dose reduction exceeds >100-fold.
Reduced photobleaching allows for brighter images at resolutions below 30 nm.

Conclusions:

DyMIN scanning offers a significant reduction in light dose for super-resolution microscopy.
This method preserves sample integrity by minimizing photobleaching.
DyMIN enables acquisition of higher quality, brighter super-resolution images with reduced exposure.