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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 developed.

Confocal Fluorescence Microscopy

Confocal Fluorescence Microscopy

Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...

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Related Experiment Video

Updated: May 27, 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

Parallelized STED fluorescence nanoscopy.

Pit Bingen¹, Matthias Reuss, Johann Engelhardt

¹German Cancer Research Center (DKFZ), Optical Nanoscopy Division, Im Neuenheimer Feld 280, 69120 Heidelberg, Germany.

|November 24, 2011

Summary

This summary is machine-generated.

We developed a parallelized STED microscope with 4 scanning beams, increasing imaging speed fourfold while retaining STED microscopy advantages. This inherently aligned system is scalable for faster super-resolution imaging.

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Last Updated: May 27, 2026

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

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Published on: March 24, 2014

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Published on: October 28, 2018

Area of Science:

Super-resolution microscopy
Optical imaging technologies
Biophysics

Background:

Stimulated Emission Depletion (STED) microscopy offers high-resolution imaging.
Achieving faster STED imaging is crucial for observing dynamic biological processes.

Purpose of the Study:

To develop a parallelized STED microscope for enhanced imaging speed.
To maintain the benefits of single-beam STED microscopy in a multi-beam setup.

Main Methods:

Implementation of a parallelized STED microscope with m=4 pairs of scanning excitation and STED beams.
Utilizing a single laser source and fiber input for inherent spatial and temporal alignment.

Main Results:

Achieved m-fold (fourfold) increase in imaging speed for a given sample area.
Maintained the advantages of single-beam scanning STED microscopy.
Demonstrated inherent spatial and temporal alignment of the parallel beams.

Conclusions:

The parallelized STED microscope significantly enhances imaging speed.
The design is scalable to higher degrees of parallelization (m) with sufficient laser power.
This technology facilitates faster super-resolution imaging of biological samples.