Recent advances on organic fluorescent probes for stimulated emission depletion (STED) microscopy

2020 ◽  
Vol 23 ◽  
Author(s):  
Ruohan Xu ◽  
Yanzi Xu ◽  
Zhi Wang ◽  
Yu Zhou ◽  
Dongfeng Dang ◽  
...  
Keyword(s):  
Fluorescent Probes ◽  
Stimulated Emission ◽  
Biological Research ◽  
Future Research ◽  
Sted Microscopy ◽  
Recent Advances ◽  
Polymer Dots ◽  
Stimulated Emission Depletion ◽  
Photo Stability

: Stimulated emission depletion (STED) microscopy has become a useful tool to visualize and dynamic monitor at an ultra-high resolution in both biological research and material science. For STED technology, fluorescent probes are irreplaceable in imaging process. Among them, organic fluorescent probes exhibit characteristics of superior photo-stability, high brightness, large Stokes' shifts and excellent biocompability, thus resulting in wide applications in STED microscopy. Based on this consideration, in this review, the recent advances on organic fluorescent probes, including typical organic fluorescent probes, aggregation-induced emission luminogens (AIEgens), polymer dots and other nanoparticles, are introduced. The applications of organic fluorescent probes in biological imaging, such as live-cell, tissue and in-vivo imaging, and also material monitoring on the nanometer scale by using STED microscopy are then included. Based on these results, the rules to design new materials for STED microscopy are provided to enhance their imaging performance and then further enrich their real-world applications in future research.

Recent advances in luminescent materials for super-resolution imaging via stimulated emission depletion nanoscopy

2021 ◽  
Author(s):  
Yanzi Xu ◽  
Ruohan Xu ◽  
Zhi Wang ◽  
Yu Zhou ◽  
Qifei Shen ◽  
...  
Keyword(s):  
Recent Progress ◽  
Super Resolution ◽  
Stimulated Emission ◽  
Luminescent Materials ◽  
New Materials ◽  
Recent Advances ◽  
Resolution Imaging ◽  
Stimulated Emission Depletion

Recent progress on STED fluorophores for super-resolution imaging and also their characteristics are outlined here, thus providing some guidelines to select proper probes and even develop new materials for super-resolution imaging via STED nanoscopy.

scholarly journals The in vivo mechanics of the magnetotactic backbone as revealed by correlative FLIM-FRET and STED microscopy

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Erika Günther ◽  
André Klauß ◽  
Mauricio Toro-Nahuelpan ◽  
Dirk Schüler ◽  
Carsten Hille ◽  
...  
Keyword(s):  
Magnetic Particles ◽  
Fluorescent Protein ◽  
Ex Vivo ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Living Cells ◽  
Stimulated Emission ◽  
Fluorescence Lifetime Imaging ◽  
Sted Microscopy

AbstractProtein interaction and protein imaging strongly benefit from the advancements in time-resolved and superresolution fluorescence microscopic techniques. However, the techniques were typically applied separately and ex vivo because of technical challenges and the absence of suitable fluorescent protein pairs. Here, we show correlative in vivo fluorescence lifetime imaging microscopy Förster resonance energy transfer (FLIM-FRET) and stimulated emission depletion (STED) microscopy to unravel protein mechanics and structure in living cells. We use magnetotactic bacteria as a model system where two proteins, MamJ and MamK, are used to assemble magnetic particles called magnetosomes. The filament polymerizes out of MamK and the magnetosomes are connected via the linker MamJ. Our system reveals that bacterial filamentous structures are more fragile than the connection of biomineralized particles to this filament. More importantly, we anticipate the technique to find wide applicability for the study and quantification of biological processes in living cells and at high resolution.

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.

scholarly journals Shedding New Lights Into STED Microscopy: Emerging Nanoprobes for Imaging

2021 ◽  
Vol 9 ◽  
Author(s):  
Yanfeng Liu ◽  
Zheng Peng ◽  
Xiao Peng ◽  
Wei Yan ◽  
Zhigang Yang ◽  
...  
Keyword(s):  
Molecular Probes ◽  
Research Field ◽  
Stimulated Emission ◽  
Light Power ◽  
Sted Microscopy ◽  
Imaging Quality ◽  
Polymer Dots ◽  
Pros And Cons

First reported in 1994, stimulated emission depletion (STED) microscopy has long been regarded as a powerful tool for real-time superresolved bioimaging . However, high STED light power (101∼3 MW/cm2) is often required to achieve significant resolution improvement, which inevitably introduces phototoxicity and severe photobleaching, damaging the imaging quality, especially for long-term cases. Recently, the employment of nanoprobes (quantum dots, upconversion nanoparticles, carbon dots, polymer dots, AIE dots, etc.) in STED imaging has brought opportunities to overcoming such long-existing issues. These nanomaterials designed for STED imaging show not only lower STED power requirements but also more efficient photoluminescence (PL) and enhanced photostability than organic molecular probes. Herein, we review the recent progress in the development of nanoprobes for STED imaging, to highlight their potential in improving the long-term imaging quality of STED microscopy and broadening its application scope. We also discuss the pros and cons for specific classes of nanoprobes for STED bioimaging in detail to provide practical references for biological researchers seeking suitable imaging kits, promoting the development of relative research field.

scholarly journals Blinking fluorescent probes for tubulin nanoscopy in living and fixed cells

2021 ◽  
Author(s):  
Ruta Gerasimaite ◽  
Jonas Bucevicius ◽  
Kamila A. Kiszka ◽  
Georgij Kostiuk ◽  
Tanja Koenen ◽  
...  
Keyword(s):  
Small Molecule ◽  
Single Molecule ◽  
Fluorescent Probes ◽  
Living Cells ◽  
Stimulated Emission ◽  
Sted Microscopy ◽  
Linker Length ◽  
Localisation Microscopy ◽  
Red Dye ◽  
Small Molecule Probe

Here we report a small molecule probe for single molecule localisation microscopy (SMLM) of tubulin in living and fixed cells. We explored a series of constructs composed of taxanes and spontaneously blinking far-red dye hydroxymethyl silicon-rhodamine (HMSiR). We found that the linker length profoundly affects the probe permeability and off-targeting. The best performing probe, HMSiR-tubulin, is composed of cabazitaxel and 6'-regioisomer of HMSiR bridged by a C6 linker. Microtubule diameters of <50 nm can be routinely measured in SMLM experiments on living and fixed cells. HMSiR-tubulin also performs well in 3D stimulated emission depletion (STED) microscopy, allowing a complementary use of both nanoscopy methods for investigating microtubule functions in living cells.

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