We combined 3D-STED microscopy and fluorescent labeling of the extracellular fluid to develop super-resolution shadow imaging (SUSHI) of brain ECS in living organotypic brain slices. SUSHI enables quantitative analysis of ECS structure and reveals dynamics on multiple scales in response to a variety of physiological stimuli.

Application in stimulated emission depletion (STED) fluorescence microscopy revealed a resolution of 50–75 nm in one- and two-color imaging

Light-Sheet Imaging. Long time-lapse imaging of live model organisms with low phototoxicity; Ultrafast volumetric imaging of cells with lattice light-sheet microscopy; Super-Resolution Imaging in Living Cells – MoNaLISA. Smart probes. 50 nm resolution at 1-2 Hz. 100 x 100 mm field of view.

Sted imaging

For single color STED imaging, SiR-tubulin or SiR-actin were added to DIV15 neurons in the BrainPhys medium at a final concentration of 1 µM during 1.5 hr at 37°C. For dual color imaging, SiR-tubulin and SiR700-actin were added to to the BrainPhys medium at 1 µM final concentration each and neurons were exposed during 1.5 hr at 37°C. Due to relatively high powers used in STED, biological samples may be affected by the illumination in the process of image acquisition. Similarly, the performance of the system may be limited by the sample itself. Optimization of the STED parameters taking into account the sample itself is therefore a complex task as there is no clear methodology that can determine the image improvement in an 2012-08-15 · Imaging and physiology data were acquired using data acquisition boards (National Instruments, Austin, TX) and modiﬁed versions of the software ScanImage (14) with code written in MATLAB (The MathWorks, Natick, MA). STED laser powers were measured at the output of the microscope objective.

A dedi-cated STED objective provides chromatic optimi- Pushing the Limits of Fluorescence Microscopy: Fluorescent Probes for Super Resolution Imaging Technologies (BioProbes 64) Beyond light's limits: Fluorescence imaging at the nanoscale—Fluorescent probes for three super-resolution modalities—STORM, SIM, and STED microscopy (BioProbes 70) Fluorescence SpectraViewer; Molecular Probes Handbook Having characterized the PSF of the system, the all-fiber STED system is used to perform STED imaging experiments.

14 Aug 2019 For STED microscopy right now, the standard of resolution is between 20 and 40 nm depending on the fluorophore, and depending on the

Stimulated Emission Depletion (STED) is a powerful microscopy technique that allows for the observation of fluorescence structure with spatial resolution below

2020-05-19 STED instruments utilize a raster-scan imaging scenario similar to a laser-scanning confocal microscope. In contrast, stochastic optical reconstruction microscopy ( STORM ), as performed using Nikon's N-STORM system, is a single-molecule approach that relies on activation of a limited subset of the overall molecular population to sequentially image and localize individual emitters on a 2020-03-01 2017-02-28 2008-12-02 2019-09-30 Using a commercial STED microscope and restricting imaging to smaller field of views allowed STED imaging at a frame rate of 0.4 to 0.8 Hz (Supplementary Videos 5 and 6). Because Nile Red can be effectively depleted using far-red depletion lasers (here λ dep = 775 nm), it can be combined with other membrane permeable labels, for example, live-cell compatible SiR-conjugates. 2018-06-22 STED Imaging Works With Quantum Dots Quantum dots survive more than 1,000 scan cycles, could be useful for time-lapse imaging by Celia Henry Arnaud June 1, 2015 | APPEARED IN VOLUME 93, ISSUE 22 We report Stimulated Emission Depletion (STED) imaging of green fluorescent nanodiamonds containing Nitrogen-Vacancy-Nitrogen (NVN) centers with a resolution of 70 nm using a commercial microscope. Nanodiamonds have been demonstrated to have the potential to be excellent cellular biomarkers thanks to their low toxicity and nonbleaching fluorescence, and are especially appealing for STED microscopy uses two laser pulses to localize fluorescence at each focal spot.

Dual-color STED imaging with spectral discrimination has been shown earlier . Since, from those experiments, our STED microscope was pre-equipped for two-color operation we were able to combine lifetime and spectral separation to perform three-channel STED imaging with resolution not fundamentally limited by diffraction.
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Together with a dedicated team in close contact with the international scientific microscopic community, we continuously improve our software, keeping it at the forefront of technology.

2021-03-15 2020-11-11 Stimulated emission depletion (STED) nanoscopy is a promising fluorescence microscopy to detect unresolvable structures at the nanoscale level and then achieve a superior imaging resolution in materials science and biological research. However, in addition to the optimization of the microscope, luminescent … With the STED technique, however, the necessary resolution can be reached, and therefore, this type of positive identification may be possible (see Fig. 2e). To improve the resolution of STED imaging even further, increased laser depletion intensity and/or smaller fluorescent probe complexes might be useful.
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Time-lapse STED imaging of the inter-mitochondrial fusion.Source: "A photostable fluorescent marker for the superresolution live imaging of the dynamic struc

Friedrich-Schiller University Jena and Leibniz Institute of 9 Jul 2020 Stimulated emission depletion (STED) microscopy provides super-resolved images by selective deactivation of fluorophores in the outer region First, we compare the STED performance imaging single molecules of several common dyes and report a novel STED dye. Then we apply STED to image 19 May 2020 Stimulated emission depletion (STED) microscopy does not suffer from these shortcomings despite having a lower axial resolution (20).

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Scientific Volume Imaging to provides reliable, high quality, easy to use image processing tools for scientists working in light microscopy. Together with a dedicated team in close contact with the international scientific microscopic community, we continuously improve our software, keeping it at the forefront of technology.

In contrast, stochastic optical reconstruction microscopy ( STORM ), as performed using Nikon's N-STORM system, is a single-molecule approach that relies on activation of a limited subset of the overall molecular population to sequentially image and localize individual emitters on a 2020-03-01 2017-02-28 2008-12-02 2019-09-30 Using a commercial STED microscope and restricting imaging to smaller field of views allowed STED imaging at a frame rate of 0.4 to 0.8 Hz (Supplementary Videos 5 and 6). Because Nile Red can be effectively depleted using far-red depletion lasers (here λ dep = 775 nm), it can be combined with other membrane permeable labels, for example, live-cell compatible SiR-conjugates. 2018-06-22 STED Imaging Works With Quantum Dots Quantum dots survive more than 1,000 scan cycles, could be useful for time-lapse imaging by Celia Henry Arnaud June 1, 2015 | APPEARED IN VOLUME 93, ISSUE 22 We report Stimulated Emission Depletion (STED) imaging of green fluorescent nanodiamonds containing Nitrogen-Vacancy-Nitrogen (NVN) centers with a resolution of 70 nm using a commercial microscope. Nanodiamonds have been demonstrated to have the potential to be excellent cellular biomarkers thanks to their low toxicity and nonbleaching fluorescence, and are especially appealing for STED microscopy uses two laser pulses to localize fluorescence at each focal spot. The first pulse is used to excite a fluorophore to its fluorescent state, and the second pulse is a modified beam used to de-excite any fluorophores surrounding the excitation focal spot.