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Related Concept Videos

Imaging Biological Samples with Optical Microscopy01:18

Imaging Biological Samples with Optical Microscopy

Optical microscopy uses optic principles to provide detailed images of samples. Antonie van Leeuwenhoek designed the first compound optical microscope in the 17th century to visualize blood cells, bacteria, and yeast cells. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes with enhanced magnification and resolution.
In optical microscopy, the specimen to be viewed is placed on a glass slide and clipped on the stage...
Simple Staining Technique01:24

Simple Staining Technique

OverviewStaining techniques in microscopy enhance the visualization of microorganisms by increasing contrast and allowing the differentiation of cellular structures. Simple staining is one of the fundamental methods used to observe the basic morphological characteristics of microorganisms, including their size, shape, and arrangement. This method relies on the application of a single dye to stain the entire cell, producing a clear contrast between the cell and the background.FixationFixation is...
Two-Dimensional Microscopy in Microbiology01:29

Two-Dimensional Microscopy in Microbiology

Two-dimensional (2D) microscopy encompasses a range of optical techniques that capture images within a single focal plane, offering detailed representations of microscopic structures. These techniques are essential in biological and medical research, enabling the visualization of cellular and subcellular structures with different levels of contrast and specificity.There are several major types of 2D microscopy, each with strengths and applications.Bright-Field MicroscopyBright-field microscopy...
Overview of Microscopy Techniques01:22

Overview of Microscopy Techniques

The early pioneers of microscopy opened a window into the invisible world of microorganisms. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes that leveraged nonvisible light, such as fluorescence microscopy that uses an ultraviolet light source and electron microscopy that uses short-wavelength electron beams. These advances significantly improved magnification, image resolution, and contrast. By comparison, the...
Super-resolution Fluorescence Microscopy01:37

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.
Overview of Electron Microscopy01:25

Overview of Electron Microscopy

The wavelengths of visible light ultimately limit the maximum theoretical resolution of images created by light microscopes. Most light microscopes can only magnify 1000X, and a few can magnify up to 1500X. Electrons, like electromagnetic radiation, can behave like waves, but with wavelengths of 0.005 nm, they produce significantly greater resolution up to 0.05 nm as compared to 500 nm for visible light. An electron microscope (EM) can create a sharp image that is magnified up to 2,000,000X.

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Related Experiment Video

Updated: Jun 5, 2026

Visualization of the Immunological Synapse by Dual Color Time-gated Stimulated Emission Depletion (STED) Nanoscopy
10:00

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

Published on: March 24, 2014

Analytical description of STED microscopy performance.

Marcel Leutenegger1, Christian Eggeling, Stefan W Hell

  • 1Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany. marcel.leutenegger@a3.epfl.ch

Optics Express
|December 18, 2010
PubMed
Summary
This summary is machine-generated.

We developed analytical equations to estimate fluorophore darkening efficiency in Stimulated Emission Depletion (STED) microscopy. This enables quick optimization of resolution and contrast for advanced imaging applications.

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Last Updated: Jun 5, 2026

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

Published on: March 24, 2014

Visualizing the Actin and Microtubule Cytoskeletons at the B-cell Immune Synapse Using Stimulated Emission Depletion (STED) Microscopy
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Visualizing the Actin and Microtubule Cytoskeletons at the B-cell Immune Synapse Using Stimulated Emission Depletion (STED) Microscopy

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Optimized Negative Staining: a High-throughput Protocol for Examining Small and Asymmetric Protein Structure by Electron Microscopy
09:37

Optimized Negative Staining: a High-throughput Protocol for Examining Small and Asymmetric Protein Structure by Electron Microscopy

Published on: August 15, 2014

Area of Science:

  • Optics and Photonics
  • Biophysical Techniques
  • Microscopy

Background:

  • Stimulated Emission Depletion (STED) microscopy overcomes the far-field optical diffraction limit.
  • STED achieves super-resolution by selectively deactivating fluorophores.
  • Efficient fluorophore deactivation is crucial for STED performance.

Purpose of the Study:

  • To develop analytical equations for estimating transient fluorophore darkening efficiency in STED.
  • To enable quick analysis and optimization of STED imaging resolution and contrast.
  • To investigate the impact of STED beam characteristics on deactivation efficiency.

Main Methods:

  • Developed analytical equations based on the spatio-temporal intensity profile of the STED beam.
  • Considered continuous wave (CW) and pulsed STED beam configurations.
  • Analyzed various pulse durations and their effect on deactivation.
  • Applied the equations to fluorescence fluctuation methods, specifically STED-Fluorescence Correlation Spectroscopy (STED-FCS).

Main Results:

  • The derived equations accurately estimate fluorophore darkening efficiency.
  • The analysis provides insights into optimizing resolution and contrast under different STED conditions.
  • The method is applicable to both CW and pulsed STED, including varying pulse durations.
  • Demonstrated utility in enhancing STED-FCS measurements.

Conclusions:

  • The developed analytical framework offers a rapid method for STED system optimization.
  • This approach facilitates improved resolution and contrast in super-resolution microscopy.
  • The findings are particularly relevant for advanced fluorescence fluctuation spectroscopy techniques like STED-FCS.