Related Experiment Video
Updated: Mar 15, 2026
10:29
Studying Dynamic Processes of Nano-sized Objects in Liquid using Scanning Transmission Electron Microscopy
Published on: February 5, 2017
13.2K
From single molecules to life: microscopy at the nanoscale
Bartosz Turkowyd1, David Virant1, Ulrike Endesfelder2
1Department of Systems and Synthetic Microbiology, Max Planck Institute for Terrestrial Microbiology and LOEWE Center for Synthetic Microbiology (SYNMIKRO), Karl-von-Frisch-Str. 16, 35043, Marburg, Germany.
Analytical and Bioanalytical Chemistry
|September 11, 2016
Summary
Super-resolution microscopy breaks the diffraction limit for nanoscale imaging of cells. This review covers stimulated-emission depletion (STED), structured illumination microscopy (SIM), and single-molecule localization microscopy (SMLM) techniques.
Area of Science:
- Biophysics
- Cell Biology
- Optical Microscopy
Background:
- The diffraction limit of light restricts resolution in conventional fluorescence microscopy to ~200 nm.
- Super-resolution microscopy techniques have revolutionized nanoscale imaging of cellular structures and processes.
- These advanced methods are crucial for modern biological research.
Purpose of the Study:
- To review the fundamental principles and practical aspects of super-resolution microscopy.
- To provide a guide for designing and implementing super-resolution experiments.
- To highlight recent advancements and future directions in the field.
Main Methods:
- Detailed theoretical overview of stimulated-emission depletion (STED) microscopy.
- Explanation of structured illumination microscopy (SIM) principles.
- Discussion of single-molecule localization microscopy (SMLM) methodologies.
Main Results:
- Comprehensive guide to experimental design, including photochemistry, labeling, and sample preparation.
- Presentation of the underlying physics and working principles of STED, SIM, and SMLM.
- Overview of practical considerations for implementing these techniques.
Conclusions:
- Super-resolution microscopy offers unprecedented nanoscale insights into cell biology.
- STED, SIM, and SMLM are key techniques for overcoming the diffraction barrier.
- The field continues to evolve with exciting applications and ongoing developments.
Keywords:
Live cell imagingPhotophysics and photochemistry of fluorophoresQuantitative cell biologySuper-resolution microscopyMore Related Videos
Related Concept Videos
01:25Overview of Electron Microscopy
16.1K
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.
16.1K
01:22Overview of Microscopy Techniques
17.6K
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...
17.6K
01:37Super-resolution Fluorescence Microscopy
14.7K
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...
14.7K
01:18Imaging Biological Samples with Optical Microscopy
12.1K
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...
In optical microscopy, the specimen to be viewed is placed on a glass slide and clipped on the stage...
12.1K
01:08Atomic Force Microscopy
4.6K
Atomic force microscopy (AFM) is a type of scanning probe microscopy that can analyze topographic details of various specimens like ceramics, glass, polymers, and biological samples. AFM offers over 1000 times more resolution than the optical imaging system. Images generated from AFM are three-dimensional surface profiles, offering an advantage over the flat, two-dimensional images from other imaging techniques.
The AFM Probe
The probe is regarded as the heart of any AFM setup and comprises the...
The AFM Probe
The probe is regarded as the heart of any AFM setup and comprises the...
4.6K
01:15Transmission Electron Microscopy
7.7K
In 1931, physicist Ernst Ruska—building on the idea that magnetic fields can direct an electron beam just as lenses can direct a beam of light in an optical microscope—developed the first prototype of the electron microscope. This development led to the development of the field of electron microscopy. In the transmission electron microscope (TEM), electrons are produced by a hot tungsten element and accelerated by a potential difference in an electron gun, which gives them up to 400...
7.7K