Super-Resolution Microscopy

Super-resolution light microscopy empowers you to study subcellular structures and dynamics with greater detail. While the spatial resolution of confocal image acquisition is doubled with the LIGHTNING technology, when using STED it can deliver insights at the nanoscale. Learn more here about Leica super-resolution methods and how they enable novel discoveries in the fields of virology, immunology, neuroscience, and cancer research.

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STED Microscopes - Stimulated Emission Depletion

About Super-Resolution

Due to the diffraction limit of light, “traditional” fluorescence microscopy techniques cannot resolve structures below 240 nm. Because biology does not stop at this scale, so-called super-resolution microscopy techniques are indispensable. LIGHTNING and STED are two compelling Leica super-resolution confocal methods for visualizing structures with greater precision and uncovering details that would otherwise not be visible.

The LIGHTNING detection concept is based on adaptive deconvolution and effectively doubles the lateral resolution to 120 nm. With LIGHTNING, you can probe specimens from microtubule dynamics to the structure of subcellular compartments.

STED nanoscopy takes super resolution to the next level, enabling that you routinely achieve lateral resolutions below 30 nm. With STED, you can dissect cellular structure and function in vivo, such as the re-organization of chromatin and nuclear pores, the finest changes in neuronal structure, intracellular transport of mitochondria and synaptic vesicles, the entry of virus particles, the colocalization of protein complexes, and more.

Frequently Asked Questions Super Resolution

Super-resolution microscopy is an optical imaging technology that overcomes the diffraction limit of light and allows the visualization of subcellular structures and dynamics in greater detail than can be achieved with conventional optical microscopy. A resolution of 30 nm is possible using STED (stimulated emission depletion) with nanoscopy. A nanoscale resolution reveals information about subcellular structures and interactions with unprecedented detail. Super-resolution microscopy enables groundbreaking discoveries in the fields of virology, immunology, neuroscience, and cancer research.

For more information about super-resolution microscopy, refer to the Science Lab articles mentioned above or go to: The Microscopy Knowledge Portal

Super-resolution microscopy can be performed in several ways. Here are the 3 more common methods offered by Leica Microsystems:

  1. The LIGHTNING detection concept enables super-resolution confocal microscopy. Unlike traditional deconvolution techniques that use a global set of parameters for the full image, it is based on an adaptive deconvolution process which uses a voxel-specific decision mask for finding the optimal deconvolution parameters . Thanks to powerful GPU computing, LIGHTNING works in near real-time for multiple colors simultaneously. In combination with a resonant scanner, it also provides a large field of view and high frame rate. For more information about LIGHTNING, refer to the following: Obtain maximum information from your specimen with LIGHTNING & Image information extraction by adaptive deconvolution
  2. The STED method along with confocal also allows super-resolution fluorescence microscopy. STED works with multiple channels and approaches isotropic super-resolution imaging in three dimensions. STED is based on the illumination of a diffraction-limited spot with a fluorophore-exciting wavelength and simultaneous illumination of a ring-shaped area with wavelengths that cause de-excitation or stimulated emission. This arrangement renders only a sub-diffraction area in the excited state and consequently allows probing with higher resolution than that of typical diffraction-limited optical microscopy. The unique TauSTED functionality from Leica Microsystems even allows for the increased STED resolution while eliminating undesired background noise .

For more information about STED, refer to: Video Talk on Super-Resolution: Overview and Stimulated Emission Depletion (STED) Microscopy & TauSTED: pushing STED beyond its limits with lifetime

Whereas super-resolution microscopy exploits visible light optics to visualize a sample, electron microscopy exploits a beam of electrons for imaging a sample. Both techniques have their pros and cons, yet only super-resolution microscopy offers the advantages of experimenting with living specimens, labeling multiple molecular targets to give them specific contrast, and convenient sample preparation.

Life Science Research

Leica Microsystems’ life science research microscopes support the imaging needs of the scientific community with advanced innovation and technical expertise for the visualization, measurement and analysis of microstructures.

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Do your research interests focus on viral infection and disease? Find out how you can gain insights into virology with solutions for imaging and sample preparation from Leica Microsystems.

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Are you working towards a better understanding of neurodegenerative diseases or studying the function of the nervous system? See how you can make breakthroughs with imaging solutions from Leica Microsystems.

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Cancer Research

Cancer is a complex and heterogeneous disease caused by cells deficient in growth regulation. Genetic and epigenetic changes in one or a group of cells disrupt normal function and result in autonomous, uncontrolled cell growth and proliferation.

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Related Articles

Read our latest articles about Super-Resolution Microscopy

The knowledge portal of Leica Microsystems offers scientific research and teaching material on the subjects of microscopy. The content is designed to support beginners, experienced practitioners and scientists alike in their everyday work and experiments.

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A Versatile Palette of Fluorescent Probes

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STED imaging of mitochondria revealed cristae dynamics during fusion. A comparison of confocal (left) and STED (right) imaging.

Mitochondrial dynamics quantitatively revealed by STED nanoscopy

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Benefits of Combining STED and Lifetime

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Effects of Clearing Media on Tissue Transparency and Shrinkage

This study comprehensively evaluates the effects of different clearing media on tissue transparency and shrinkage by comparing freshly dissected dipteran fly brains with their cleared equivalents.…
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Visualization of Submitochondrial Protein Distributions

By allowing visualization of submitochondrial protein distributions, fluorescence nanoscopy offers significant advantages for understanding cell death control. How cells manage, and control…
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DNA Replication in Cancer Cells

DNA synthesis can be impeded by collisions between the DNA replication machinery and co-transcriptional R-loops leading to a major source of genomic instability in cancer cells. In this paper we…
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Super-Resolution Microscopy Image Gallery

Due to the diffraction limit of light, traditional confocal microscopy cannot resolve structures below ~240 nm. Super-resolution microscopy techniques, such as STED, PALM or STORM or some…
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Regulators of Actin Cytoskeletal Regulation and Cell Migration in Human NK Cells

Dr. Mace will describe new advances in our understanding of the regulation of human NK cell actin cytoskeletal remodeling in cell migration and immune synapse formation derived from confocal and…
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Benefits of TauContrast to Image Complex Samples

In this interview, Dr. Timo Zimmermann talks about his experience with the application of TauSense tools and their potential for the investigation of demanding samples such as thick samples or…
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Microscopy in Virology

The coronavirus SARS-CoV-2, causing the Covid-19 disease effects our world in all aspects. Research to find immunization and treatment methods, in other words to fight this virus, gained highest…
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New Light Shed on the Nanodomain Organization of the Endoplasmic Reticulum (ER)

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Cover glomerulus

The Guide to STED Sample Preparation

This guide is intended to help users optimize sample preparation for stimulated emission depletion (STED) nanoscopy, specifically when using the TCS SP8 STED 3X nanoscope from Leica Microsystems. It…
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Extending Nanoscopy Possibilities with STED and exchangeable fluorophores

When it comes to STED Nanoscopy, keeping high signal-to-noise is key to achieve the best possible resolution in fixed and living cells. This can be challenging in the case of experiments in 3D and/or…
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See More Than Just Your Image

Despite the emergence of new imaging methods in recent years, true 3D resolution is still achieved by Confocal Laser Scanning Microscopy (CLSM). Through a combination of novel, extremely fast scanning…
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Simultaneously Measuring Image Features and Resolution in Live-Cell STED Images

Reliable interpretation and quantification of cellular features in fluorescence microscopy requires an accurate estimate of microscope resolution. This is typically obtained by measuring the image of…
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Observing Malaria Infection at the Right Spot in the Human Host

Malaria is a life-threatening disease transmitted through the bites of mosquitoes infected with protozoan parasites. The most common and dangerous type of malaria is caused by the parasite Plasmodium…
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Super-resolved STED spectroscopy

Molecular interactions are key in cellular signalling. They are often ruled or rendered by the mobility of the involved molecules.
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STED Nanoscopy at the forefront of cancer research

Alison Dun is the postdoctoral facility manager for the Edinburgh Super-Resolution Imaging Consortium (ESRIC), Heriot-Watt University, Edinburgh, UK. She has used a large range of microscope…
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