Giovanna Coceano - Combined STED nanoscopy and enzymatic labeling to study the fine organization and dynamics of neuronal organelles

Stimulated Emission Depletion microscopy, or STED nanoscopy, is a technique that uses the non-linear de-excitation of fluorescent dyes to overcome the

It discusses the practical aspects in performing optimal STED experiment. Finally, it discusses the applications of STED nanoscopy in biological systems with special relevance to membrane biology. 2. Sr- y Fluorescence microscopy has become an inevitable tool in model cell biology studies.

Here we present a dual-color STED nanoscope utilizing two orthogonally crossed Double-helix enhanced axial localization in STED nanoscopy G. P. J. Laporte,1,* D. B. Conkey,2 A. Vasdekis,3 R. Piestun,2 and D. Psaltis1 1Laboratory of Optics, School of Engineering, École Poly technique Fédérale de Lausanne (EPFL), Station 17, 1015 dual-color STED nanoscopy (Fig. 1) and molecular diffu-sion quantiﬁcation down to ~20 nm in (living) cells. The presented dual-channel STED microscope utilizes a single ﬁber laser providing a 20-MHz train of 775 nm wavelength pulses of 1.2-ns duration. This compact laser source en-ables STED on ﬂuorophores emitting in the orange to red range. STED Nanoscopy of Interfaces and Interactions between Nanostructure Arrays and Living Cells.

The biological structures that need to be examined to understand diseases, such as cancer, is on such a small scale, so small we still cannot truly know and see with our own eyes what is happening inside cells. However STED nanoscopy allows us to get closer to visualize these than ever before, revealing structures inside cells not previously visible. As already mentioned in Sect.

2018-11-12 · a STED nanoscopy of a HeLa cell expressing VIM-SNIFP. Large image: magnification of the region indicated in the inset showing the STED image of a whole cell (see also Supplementary Fig. 5 ).

By: Ilaria Testa, Cellular Biophysics, KTH/SciLife. Title: "STED/RESOLFT optical nanoscopy: applications to the life sciences". Choosing dyes for cw-STED nanoscopy using self-assembled nanorulers. S Beater, P Holzmeister, E Pibiri, B Lalkens, P Tinnefeld.

As already mentioned in Sect. 1.2.1, STED nanoscopy is based on the idea of limiting the ability of molecules to fluoresce in the immediate vicinity of the geometric focus. Since the fluorescence is inhibited by the process of stimulated emission, it is necessary to shape the intensity distribution of the STED light such that it is zero in the geometric focus.

The stimulated emission is induced by the STED light at a wavelength that is typically at the red edge of the emission spectrum. STED nanoscopy: imaging beyond the diffraction limit of light, 23rd Dubai International Pharmaceuticals and Technologies Conference and Exhibition DUPHAT 2018, Dubai (02.2018)J.-R. Krüger, B. Vinçon, C. Geisler, A. Egner: Learn more: https://www.ibiology.org/talks/stimulated-emission-depletion/Historically, light microscopy has been limited in its ability to resolve closely sp 2012-09-28 · For STED microscopy and the corresponding confocal microscopy, either a stage scanning STED microscope (Exc. 635 nm; STED 775 nm/ 20 MHz/ 1 ns pulses from frequency-doubled fiber laser; Detection range 670/40 nm) or a beam scanning STED microscope (Exc. 640 nm; STED 760 nm/ 76 MHz/ ∼ 200 ps pulses up-chirped from 100 fs mode-locked Ti: Sapphire laser; Detection range 670/40 nm) was used. Aug 9, 2016 Commonly, in stimulated emission depletion (STED) fluorescence nanoscopy, light of a wavelength located at the red tail of the emission Jan 27, 2020 In this study, we attempted to use commercially available CdSe@ZnS QDs with green emission (QD526) as a probe in STED nanoscopy. Jan 18, 2013 For this purpose, STED microscopy (or nanoscopy) uses stimulated emission to inhibit fluorescence emission at predefined sample coordinates Jun 19, 2019 We coupled a stimulated emission depletion (STED) microscope with an atomic force microscope to investigate the formation of amyloid Jun 1, 2020 Abstract Super‐resolution microscopy techniques can provide answers to still pending questions on prokaryotic organisms but are yet to be the proof-of-concept to easily applicable tools in fluorescence microscopy.

Krüger, B. Vinçon, C. Geisler, A. Egner: Learn more: https://www.ibiology.org/talks/stimulated-emission-depletion/Historically, light microscopy has been limited in its ability to resolve closely sp 2012-09-28 · For STED microscopy and the corresponding confocal microscopy, either a stage scanning STED microscope (Exc. 635 nm; STED 775 nm/ 20 MHz/ 1 ns pulses from frequency-doubled fiber laser; Detection range 670/40 nm) or a beam scanning STED microscope (Exc. 640 nm; STED 760 nm/ 76 MHz/ ∼ 200 ps pulses up-chirped from 100 fs mode-locked Ti: Sapphire laser; Detection range 670/40 nm) was used. Aug 9, 2016 Commonly, in stimulated emission depletion (STED) fluorescence nanoscopy, light of a wavelength located at the red tail of the emission Jan 27, 2020 In this study, we attempted to use commercially available CdSe@ZnS QDs with green emission (QD526) as a probe in STED nanoscopy.
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This fact has been recently used to substantially improve the effective spatial resolution in STED nanoscopy using time-gated detection, pulsed excitation and continuous wave (CW) STED beams. We present a AFM-STED correlative nanoscopy reveals a dark side in fluorescence microscopy imaging By Michela Cosentino , Claudio Canale , Paolo Bianchini , Alberto Diaspro Science Advances 19 Jun 2019 : eaav8062 The low saturation intensity and high photostability of MitoESq-635 make it ideal for long-term, high-resolution (stimulated emission depletion) STED nanoscopy. We performed time-lapse imaging of the mitochondrial inner membrane over 50 min (3.9 s per frame, with 71.5 s dark recovery) in living HeLa cells with a resolution of 35.2 nm.

Herein, with the highly emissive Adaptive-illumination STED nanoscopy The concepts called STED/RESOLFT superresolve features by a light-driven transfer of closely packed molecules between two different states, typically a nonfluorescent "off" state and a fluorescent "on" state at well-defined coordinates on subdiffraction scales.
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Multi-Color 2D and 3D STED microscopy ; Image acquisition strategies for live-cell STED ; STED Fluorescence Correlation Spectroscopy (STED-FCS) Learning Outcomes. At the end of the course, the participants will be able to design experiments using STED microscopy, prepare high-quality microscopy samples and acquire state-of-the-art nanoscopy images.

Jul 27, 2015 angles and beam distortions render the system complex16–18. ultrafast, temporally stochastic sted nanoscopy of millisecond dynamics.

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Double-helix enhanced axial localization in STED nanoscopy G. P. J. Laporte,1,* D. B. Conkey,2 A. Vasdekis,3 R. Piestun,2 and D. Psaltis1 1Laboratory of Optics, School of Engineering, École Poly technique Fédérale de Lausanne (EPFL), Station 17, 1015

This compact laser source en-ables STED on ﬂuorophores emitting in the orange to red range.