An indirectly stimulated emission depletion method for STED microscopy

2019 ◽  
Vol 52 (41) ◽  
pp. 415108
Author(s):  
Zhi-Jun Luo ◽  
Ya-Nan Liu ◽  
Meng-Lin Chen ◽  
Zong-Song Gan ◽  
Chang-Sheng Xie
Keyword(s):  
Stimulated Emission ◽  
Sted Microscopy ◽  
Stimulated Emission Depletion

scholarly journals Precision targeted ruthenium(ii) luminophores; highly effective probes for cell imaging by stimulated emission depletion (STED) microscopy

2016 ◽  
Vol 7 (10) ◽  
pp. 6551-6562 ◽  
Author(s):  
Aisling Byrne ◽  
Christopher S. Burke ◽  
Tia E. Keyes
Keyword(s):  
Cell Imaging ◽  
Stimulated Emission ◽  
Cell Organelles ◽  
Polypyridyl Complexes ◽  
Sted Microscopy ◽  
Highly Effective ◽  
Stimulated Emission Depletion ◽  
Peptide Targeting

Using precision peptide targeting to discrete cell organelles, it is demonstrated that Ru(ii) polypyridyl complexes are highly effective probes for stimulated emission depletion microscopy.

scholarly journals Expansion stimulated emission depletion microscopy (ExSTED)

2018 ◽  
Author(s):  
Mengfei Gao ◽  
Riccardo Maraspini ◽  
Oliver Beutel ◽  
Amin Zehtabian ◽  
Britta Eickholt ◽  
...  
Keyword(s):  
Excited State ◽  
Optical Resolution ◽  
Super Resolution ◽  
Stimulated Emission ◽  
High Fidelity ◽  
Sted Microscopy ◽  
Structural Details ◽  
Stimulated Emission Depletion ◽  
Conventional Microscopy ◽  
Super Resolution Microscopy

AbstractStimulated emission depletion (STED) microscopy is routinely used to resolve the ultra-structure of cells with a ∼10-fold higher resolution compared to diffraction limited imaging. While STED microscopy is based on preparing the excited state of fluorescent probes with light, the recently developed expansion microscopy (ExM) provides sub-diffraction resolution by physically enlarging the sample before microscopy. Expansion of fixed cells by crosslinking and swelling of hydrogels easily enlarges the sample ∼4-fold and hence increases the effective optical resolution by this factor. To overcome the current limits of these complimentary approaches, we here combined ExM with STED (ExSTED) and demonstrate an increase in resolution of up to 30-fold compared to conventional microscopy (<10 nm lateral and ∼50 nm isotropic). While the increase in resolution is straight forward, we found that high fidelity labelling via multi-epitopes is required to obtain emitter densities that allow to resolve ultra-structural details with ExSTED. Our work provides a robust template for super resolution microscopy of entire cells in the ten nanometer range.

Integrated dual-color stimulated emission depletion (STED) microscopy and fluorescence emission difference (FED) microscopy

2018 ◽  
Vol 423 ◽  
pp. 167-174 ◽  
Author(s):  
Wensheng Wang ◽  
Guangyuan Zhao ◽  
Cuifang Kuang ◽  
Liang Xu ◽  
Shaocong Liu ◽  
...  
Keyword(s):  
Fluorescence Emission ◽  
Stimulated Emission ◽  
Sted Microscopy ◽  
Dual Color ◽  
Stimulated Emission Depletion

STED microscopy—super-resolution bio-imaging utilizing a stimulated emission depletion

Microscopy ◽  
2015 ◽  
Vol 64 (4) ◽  
pp. 227-236 ◽  
Author(s):  
Kohei Otomo ◽  
Terumasa Hibi ◽  
Yuichi Kozawa ◽  
Tomomi Nemoto
Keyword(s):  
Super Resolution ◽  
Stimulated Emission ◽  
Sted Microscopy ◽  
Bio Imaging ◽  
Stimulated Emission Depletion

scholarly journals Synergic Combination of Stimulated Emission Depletion Microscopy with Image Scanning Microscopy to Reduce Light Dosage

2019 ◽  
Author(s):  
Giorgio Tortarolo ◽  
Marco Castello ◽  
Sami Koho ◽  
Giuseppe Vicidomini
Keyword(s):  
Single Photon ◽  
Simulated Data ◽  
Live Cell ◽  
Stimulated Emission ◽  
Beam Intensity ◽  
Scanning Microscopy ◽  
Image Scanning ◽  
Sted Microscopy ◽  
Stimulated Emission Depletion

AbstractStimulated emission depletion (STED) microscopy is one of the most influential nanoscopy techniques; by increasing the STED beam intensity, it theoretically improves the spatial resolution to any desired value. However, the higher is the dose of stimulating photons, the stronger are the photo-bleaching and photo-toxicity effects, which potentially compromise live-cell and long-term imaging. For this reason the scientific community is looking for strategies to reduce the STED beam intensity needed to achieve a target resolution. Here, we show how the combination of STED microscopy with image scanning microscopy (ISM) meets this request. In particular, we introduce a new STED-ISM architecture – based on our recent single-photon-avalanche-diode (SPAD) detector array – which allows covering the near-diffraction limit resolution range with reduced STED beam intensity. We demonstrate this ability both with simulated data and in live-cell experiments. Because of (i) the minimal changes in the optical architecture of the typical point-scanning STED microscope; (ii) the parameter-free, robust and real-time pixel-reassignment method to obtain the STED-ISM image; (iii) the compatibility with all the recent progresses in STED microscopy, we envisage a natural and rapid upgrade of any STED microscope to the proposed STED-ISM architecture.

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