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Site localisation on bacterial surface proteins using super

Step 2: Choice of fluorophores There is a wide range of fluorophores performing well with STED imaging and experiments with low labeling densities designed to stay close to physiological conditions are typical. At the same time, the dead-time compensation of DynamicPLUS allows crisp images of high-signal samples, such as bright confocal images. STEDYFOCUS Module. 2020-04-07 As the spatial resolution of STED imaging correlates with an increase in STED light intensity, photodegradation of fluorescent dyes becomes a serious issue. Representative photoresistant fluorescent dyes such as Alexa Fluor® 488 and ATTO 488 are also known to encounter photodegradation in STED microscopy, making it difficult to conduct continuous live-imaging of biological systems while Super-resolution imaging using the principles of stimulated emission depletion (STED) microscopy requires collinear excitation of a sample with a Gaussian-shaped excitation beam and a donut-shaped depletion beam whose spin (polarization) and orbital angular momentum (OAM) signs are aligned.

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This ultimately enables imaging of triple color labeling with the 592 STED line (e.g. ST AR 440SX, Oregon Green 488 and Alexa Fluor 532). Gated STED triple color imaging with three standard dyes is achieved with the 660 STED laser (e.g. Alexa 488, Alexa Fluor 532 and TMR). STED microscopy delivers the most reliable super-resolved Then, the cells were washed 3 time with fresh medium to remove the free dye, and kept in DMEM+ for imaging. The Leica TCS SP8 STED 3× system with 2 CW lasers at 592 and 660 nm and a pulsed laser at 775 nm for the depletion was used for STED imaging. An HyD detector and an STED WHITE objective (100×/1.40 OIL or 93×/1.30 GLYC) were employed.

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STED Microscopes STELLARIS STED & STELLARIS 8 STED Our STED technology joins the STELLARIS platform to provide you the fastest way of imaging beyond the diffraction limit. Obtain cutting-edge nanoscopy results in no time with astounding image quality and resolution, while protecting your sample.

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Switch on the PC Microscope, Scanner Power and Laser Power buttons and turn the Laser Emission key from Off to On. Wait for the computer to boot up, login to the account and then turn on the depletion lasers. 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. Time lapse imaging; STED … 2019-06-01 Dual Color STED Imaging Leica TCS STED CW The Leica TCS STED CW is compatible with a variety of common fl uorophores such as Alexa 488, FITC, Oregon Green 488, and also genetically encoded markers like eYFP, Venus, and Citrin.

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. Our STED technology joins the STELLARIS platform to provide you the fastest way of imaging beyond the diffraction limit. Obtain cutting-edge nanoscopy results in no time with astounding image quality and resolution, while protecting your sample. Introductory guide to STED super-resolution imaging using the Leica Microsystems TCS SP8 STED 3X. Microscope start-up.
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bioRxiv @biorxivpreprint. Tiled STED Imaging of Extended Sample Regions http://biorxiv.org/cgi/content/short/789487v1 … #bioRxiv.

In specimens eGFP can yield good results. 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. The beam splitter in the system is replaced by a custom-made 2 mm thick dichroic mirror (ZT670dcrb, Chroma, USA).
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Welcome to Swedish Bioimaging 4th National Meeting

Structure and Regulation of CaMKII. M Lemieux, P De Koninck  Long-term molecular nanoscale imaging of neuronal function Ensemble approaches (including SSIM and STED) are able to record faster, but with poorer  Super resolution microscopy techniques such as stimulated emission depletion microscopy (STED) improves imaging resolution compared to  cellular biophysicsMicroscopyNephrologyneurosciencesuper-resolution in the neuron with STED and PALM microscopy and by demonstrating its functional  composite material for concurrent drug delivery and biomedical imaging.