scholarly journals Can Super-Resolution Microscopy Become a Standard Characterization Technique for Material Chemistry?

2021 ◽  
Author(s):  
Shikha Dhiman ◽  
Teodora Andrian ◽  
Beatriz Santiago ◽  
Marrit Tholen ◽  
Yuyang Wang ◽  
...  
Keyword(s):  
Super Resolution ◽  
Analytical Techniques ◽  
Material Chemistry ◽  
Characterization Technique ◽  
Super Resolution Microscopy ◽  
Synthesized Materials

The characterization of newly synthesized materials is a cornerstone of all chemistry and nanotechnology laboratories. For this purpose, a wide array of analytical techniques have been standardized and are used...

scholarly journals Characterization of Plasma Membrane Ceramides by Super-Resolution Microscopy

2017 ◽  
Vol 56 (22) ◽  
pp. 6131-6135 ◽  
Author(s):  
Anne Burgert ◽  
Jan Schlegel ◽  
Jérôme Bécam ◽  
Sören Doose ◽  
Erhard Bieberich ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Super Resolution ◽  
Super Resolution Microscopy

scholarly journals Experimental characterization of 3D localization techniques for particle-tracking and super-resolution microscopy

2009 ◽  
Vol 17 (10) ◽  
pp. 8264 ◽  
Author(s):  
Michael J. Mlodzianoski ◽  
Manuel F. Juette ◽  
Glen L. Beane ◽  
Joerg Bewersdorf
Keyword(s):  
Particle Tracking ◽  
Super Resolution ◽  
Experimental Characterization ◽  
3D Localization ◽  
Super Resolution Microscopy

scholarly journals Characterization of Ire1 Interactions and Dynamics with Quantitative Super-Resolution Microscopy

2017 ◽  
Vol 112 (3) ◽  
pp. 148a
Author(s):  
Elizabeth M. Smith ◽  
Ragnar Stefansson ◽  
Maria Paz Ramirez Lopez ◽  
Elias M. Puchner
Keyword(s):  
Super Resolution ◽  
Super Resolution Microscopy

scholarly journals Surface marker expression in small and medium/large mesenchymal stromal cell-derived extracellular vesicles in naive or apoptotic condition using orthogonal techniques

2021 ◽  
Author(s):  
Renata Skovronova ◽  
Cristina Grange ◽  
Veronica Dimuccio ◽  
Maria Chiara Deregibus ◽  
Giovanni Camussi ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Extracellular Vesicles ◽  
Super Resolution ◽  
Analytical Techniques ◽  
Surface Marker ◽  
Bulk Analysis ◽  
Surface Marker Expression ◽  
Marker Expression ◽  
Super Resolution Microscopy ◽  
Single Vesicle

Extracellular vesicles released by mesenchymal stromal cells (MSC EVs) are a promising resource for regenerative medicine. In particular, small MSC EVs represent the active EV fraction for therapeutic applications. A bulk analysis is applied to characterize MSC EVs identity and purity, coupled with the assessment of single EV morphology, size and integrity using electron microscopy. We here applied different orthogonal methods to provide a quantitative analysis of size and surface marker expression in medium/large and small fractions, namely 10k and 100k fractions, of MSC EVs obtained by sequential ultracentrifugations. Bone marrow, adipose tissue, and umbilical cord MSC EVs were compared, in naive and apoptotic conditions. The 100k EV size <100 nm, as detected by electron microscopy, was confirmed by super-resolution microscopy and ExoView. Quantitative single vesicle imaging using super-resolution microscopy revealed heterogeneous patterns of tetraspanin expressions, being all MSC EV fractions single, double and triple positive, in variable proportions, for CD63, CD81 and CD9. Moreover, ExoView analysis allowed a comparative multiplex screening of single MSC EV tetraspanin and mesenchymal marker levels. Finally, a semiquantitative bead based cytofluorimetric analysis showed the segregation of immunological and pro-coagulative markers on the 10k MSC EV fraction. Apoptotic MSC EVs were released in higher number, without significant differences from the naive fractions in surface marker expression. These results indicate that a consistent profile of MSC EV fractions among the different MSC sources, and a safer profile of the 100k MSC EV population for clinical application. Finally, our study identified suitable applications for different EV analytical techniques.

scholarly journals Towards a Quantitative Single Particle Characterization by Super Resolution Microscopy: From Virus Structures to Antivirals Design

2021 ◽  
Vol 9 ◽  
Author(s):  
Maria Arista-Romero ◽  
Silvia Pujals ◽  
Lorenzo Albertazzi
Keyword(s):  
Viral Infection ◽  
Fluorescent Dyes ◽  
Super Resolution ◽  
Particle Characterization ◽  
Emerging Viruses ◽  
Potential Threat ◽  
The Right ◽  
Super Resolution Microscopy ◽  
Selection Of

In the last year the COVID19 pandemic clearly illustrated the potential threat that viruses pose to our society. The characterization of viral structures and the identification of key proteins involved in each step of the cycle of infection are crucial to develop treatments. However, the small size of viruses, invisible under conventional fluorescence microscopy, make it difficult to study the organization of protein clusters within the viral particle. The applications of super-resolution microscopy have skyrocketed in the last years, converting this group into one of the leading techniques to characterize viruses and study the viral infection in cells, breaking the diffraction limit by achieving resolutions up to 10 nm using conventional probes such as fluorescent dyes and proteins. There are several super-resolution methods available and the selection of the right one it is crucial to study in detail all the steps involved in the viral infection, quantifying and creating models of infection for relevant viruses such as HIV-1, Influenza, herpesvirus or SARS-CoV-1. Here we review the use of super-resolution microscopy (SRM) to study all steps involved in the viral infection and antiviral design. In light of the threat of new viruses, these studies could inspire future assays to unveil the viral mechanism of emerging viruses and further develop successful antivirals against them.

scholarly journals Surface Marker Expression in Small and Medium/Large Mesenchymal Stromal Cell-Derived Extracellular Vesicles in Naive or Apoptotic Condition Using Orthogonal Techniques

Cells ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2948
Author(s):  
Renata Skovronova ◽  
Cristina Grange ◽  
Veronica Dimuccio ◽  
Maria Chiara Deregibus ◽  
Giovanni Camussi ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Extracellular Vesicles ◽  
Super Resolution ◽  
Analytical Techniques ◽  
Surface Marker ◽  
Bulk Analysis ◽  
Surface Marker Expression ◽  
Marker Expression ◽  
Super Resolution Microscopy ◽  
Different Sources

Extracellular vesicles released by mesenchymal stromal cells (MSC-EVs) are a promising resource for regenerative medicine. Small MSC-EVs represent the active EV fraction. A bulk analysis was applied to characterise MSC-EVs’ identity and purity, with the assessment of single EV morphology, size and integrity using electron microscopy. We applied different methods to quantitatively analyse the size and surface marker expression in medium/large and small fractions, namely 10k and 100k fractions, of MSC-EVs obtained using sequential ultracentrifugation. Bone marrow, adipose tissue and umbilical cord MSC-EVs were compared in naive and apoptotic conditions. As detected by electron microscopy, the 100k EV size < 100 nm was confirmed by super-resolution microscopy and ExoView. Single-vesicle imaging using super-resolution microscopy revealed heterogeneous patterns of tetraspanins. ExoView allowed a comparative screening of single MSC-EV tetraspanin and mesenchymal markers. A semiquantitative bead-based cytofluorimetric analysis showed the segregation of immunological and pro-coagulative markers on the 10k MSC-EVs. Apoptotic MSC-EVs were released in higher numbers, without significant differences in the naive fractions in surface marker expression. These results show a consistent profile of MSC-EV fractions among the different sources and a safer profile of the 100k MSC-EV population for clinical application. Our study identified suitable applications for EV analytical techniques.

scholarly journals Active diffusion and advection in theDrosophilaooplasm result from the interplay of the actin and microtubule cytoskeletons

2017 ◽  
Author(s):  
Maik Drechsler ◽  
Fabio Giavazzi ◽  
Roberto Cerbino ◽  
Isabel M. Palacios
Keyword(s):  
Actin Cytoskeleton ◽  
Super Resolution ◽  
Noisy Environment ◽  
Directed Motion ◽  
Contrast Mechanisms ◽  
Active Processes ◽  
Super Resolution Microscopy ◽  
Random Diffusion ◽  
Dynamic Link

ABSTRACTTransport in cells occurs via a delicate interplay of passive and active processes, including diffusion, directed transport and advection. Despite progresses in super-resolution microscopy, discriminating and quantifying these processes is a challenge, requiring tracking of rapidly moving, sub-diffraction objects in a crowded, noisy environment. Here we use Differential Dynamic Microscopy with different contrast mechanisms to provide a thorough characterization of the dynamics in theDrosophilaoocyte. We study the movement of vesicles and the elusive motion of a cytoplasmic F-actin mesh, a known regulator of cytoplasmic flows. We find that cytoplasmic motility constitutes a combination of directed motion and random diffusion. While advection is mainly attributed to microtubules, we find that active diffusion is driven by the actin cytoskeleton, although it is also enhanced by the flow. We also find that an important dynamic link exists between vesicles and cytoplasmic F-actin motion, as recently suggested in mouse oocytes.

scholarly journals Characterization of neuronal synaptopodin reveals a myosin V-dependent mechanism of synaptopodin clustering at the post-synaptic sites

2019 ◽  
Author(s):  
Judit Gonzalez-Gallego ◽  
Anja Konietzny ◽  
Alberto Perez-Alvarez ◽  
Julia Baer ◽  
Urban Maier ◽  
...  
Keyword(s):  
Dendritic Spines ◽  
Pyramidal Neurons ◽  
Super Resolution ◽  
Myosin V ◽  
Interaction Partners ◽  
Spine Apparatus ◽  
Tight Association ◽  
Super Resolution Microscopy ◽  
Dependent Mechanism

The spine apparatus (SA) is an endoplasmic reticulum-related organelle which is present in a subset of dendritic spines in cortical and pyramidal neurons. The synaptopodin protein localizes between the stacks of the spine apparatus and is essential for the formation of this unique organelle. Although several studies have demonstrated the significance of the SA and synaptopodin in calcium homeostasis and plasticity of dendritic spines, it is still unclear what factors contribute to its stability at the synapse and whether the SA is locally formed or it is actively delivered to the spines. In this study we show that synaptopodin clusters are stable at their locations. We found no evidence of active microtubule-based transport for synaptopodin. Instead new clusters were emerging in the spines, which we interpret as the SA being assembled on-site. Furthermore, using super-resolution microscopy we show a tight association of synaptopodin with actin filaments. We identify the actin-based motor proteins myosin V and VI as novel interaction partners of synaptopodin and demonstrate that myosin V is important for the formation and/or maintenance of the SA.

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