Stimulated Emission Depletion
Stimulated Emission Depletion - Related Videos
10:00
Visualization of the Immunological Synapse by Dual Color Time-gated Stimulated Emission Depletion (STED) Nanoscopy
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Cited by 12 •
2014
Here we illustrate the protocol for imaging by two-color STED nanoscopy the cytotoxic immune synapse of NK cells recapitulated on glass. Using this method we obtain sub-100 nm resolution of synapse proteins and the cytoskeleton.
11:00
Visualizing the Actin and Microtubule Cytoskeletons at the B-cell Immune Synapse Using Stimulated Emission Depletion (STED) Microscopy
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Cited by 10 •
2018
We present a protocol for using STED microscopy to simultaneously image actin structures, microtubules, and microtubule plus-end binding proteins in B cells that have spread on coverslips coated with antibodies to the B-cell receptor, a model for the initial phase of immune synapse formation.
06:17
Depletion of Specific Cell Populations by Complement Depletion
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Cited by 15 •
2010
To effectively study the function of immune cell populations their purification is often required. Complement depletion is a fast and inexpensive technique for the isolation of immune cell populations with high purity.
08:18
Design and Use of a Full Flow Sampling System (FFS) for the Quantification of Methane Emissions
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Cited by 13 •
2016
We have designed, developed, and implemented a novel full flow sampling system (FFS) for quantification of methane emissions and greenhouse gases from across the natural gas supply chain.
02:39
Emission Spectra
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2020
When solids, liquids, or condensed gases are heated sufficiently, they radiate some of the excess energy as light. Photons produced in this manner have a range of energies, and thereby produce a continuous spectrum in which an unbroken series of wavelengths is present. In contrast to continuous spectra, light can also occur as discrete or line spectra having very narrow linewidths interspersed throughout the spectral regions. Exciting a gas at low partial pressure using an electrical current,...
FAQs
The excitation beam energizes fluorophores, while the doughnut-shaped depletion beam drives stimulated emission around the focal center. This selectively suppresses fluorescence in the surrounding area, leaving signal from the central subdiffraction-sized region. Separating these beam functions enables spatial discrimination beyond what conventional diffraction-limited imaging can provide.
The doughnut-shaped beam surrounds the focal center with depletion light while leaving the center comparatively unsuppressed. As a result, fluorescence is removed from the region around the focus rather than from the entire illuminated area. This spatial pattern confines detectable signal and allows nanoscale features to be distinguished within biological specimens.
Conventional light microscopy cannot resolve some nanoscale structures because of the diffraction limit. Stimulated Emission Depletion restricts detectable fluorescence to a much smaller central region, improving spatial resolution. This difference makes it possible to examine protein distributions, synaptic features, organelles, and other cellular details that conventional imaging cannot separate.
First, fluorophores in the specimen are energized with a focused excitation beam. The depletion beam then suppresses fluorescence around the focal center. That confined fluorescent spot is scanned across the specimen, and the resulting position-dependent signal produces an image. The scan allows nanoscale structural information to be mapped across biological samples.
In biological research, Stimulated Emission Depletion can reveal protein distributions, organelles, synaptic features, and dynamic cellular processes. Its improved spatial resolution helps investigators study cellular architecture and molecular organization at nanoscale dimensions. The technique is therefore useful when the arrangement or behavior of fluorescently labeled features cannot be distinguished with conventional light microscopy.
Photobleaching and sample-specific labeling are important considerations when planning an experiment. The sample must provide fluorescently labeled features that can be examined with the excitation and depletion beams, while photobleaching may affect observations during imaging. These factors influence how effectively cellular structures or dynamic processes can be visualized.