microscopy techniques
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2022 ◽  
Vol 13 (1) ◽  
Author(s):  
Aleksandra Arsić ◽  
Cathleen Hagemann ◽  
Nevena Stajković ◽  
Timm Schubert ◽  
Ivana Nikić-Spiegel

AbstractModern light microscopy, including super-resolution techniques, has brought about a demand for small labeling tags that bring the fluorophore closer to the target. This challenge can be addressed by labeling unnatural amino acids (UAAs) with bioorthogonal click chemistry. The minimal size of the UAA and the possibility to couple the fluorophores directly to the protein of interest with single-residue precision in living cells make click labeling unique. Here, we establish click labeling in living primary neurons and use it for fixed-cell, live-cell, dual-color pulse–chase, and super-resolution microscopy of neurofilament light chain (NFL). We also show that click labeling can be combined with CRISPR/Cas9 genome engineering for tagging endogenous NFL. Due to its versatile nature and compatibility with advanced multicolor microscopy techniques, we anticipate that click labeling will contribute to novel discoveries in the neurobiology field.


2022 ◽  
Author(s):  
Gustave Ronteix ◽  
Valentin Bonnet ◽  
Sebastien Sart ◽  
Jeremie Sobel ◽  
Elric Esposito ◽  
...  

Microscopy techniques and image segmentation algorithms have improved dramatically this decade, leading to an ever increasing amount of biological images and a greater reliance on imaging to investigate biological questions. This has created a need for methods to extract the relevant information on the behaviors of cells and their interactions, while reducing the amount of computing power required to organize this information. This task can be performed by using a network representation in which the cells and their properties are encoded in the nodes, while the neighborhood interactions are encoded by the links. Here we introduce Griottes, an open-source tool to build the "network twin" of 2D and 3D tissues from segmented microscopy images. We show how the library can provide a wide range of biologically relevant metrics on individual cells and their neighborhoods, with the objective of providing multi-scale biological insights. The library's capacities are demonstrated on different image and data types. This library is provided as an open-source tool that can be integrated into common image analysis workflows to increase their capacities.


Diagnostics ◽  
2022 ◽  
Vol 12 (1) ◽  
pp. 192
Author(s):  
Krzysztof Lukaszuk ◽  
Grzegorz Jakiel ◽  
Izabela Wocławek Potocka ◽  
Jolanta Kiewisz ◽  
Jolanta Olszewska ◽  
...  

Intracytoplasmic sperm injection (ICSI) is a widely used and accepted treatment of choice for oocyte fertilization. However, the quality of sperm selection depends on the accurate visualization of the morphology, which can be achieved with a high image resolution. We aim to correct the conviction, shown in a myriad of publications, that an ultra-high magnification in the range of 6000×–10,000× can be achieved with an optical microscope. The goal of observing sperm under the microscope is not to simply get a larger image, but rather to obtain more detail—therefore, we indicate that the optical system’s resolution is what should be primarily considered. We provide specific microscope system setup recommendations sufficient for most clinical cases that are based on our experience showing that the optical resolution of 0.5 μm allows appropriate visualization of sperm defects. Last but not least, we suggest that mixed research results regarding the clinical value of IMSI, comparing to ICSI, can stem from a lack of standardization of microscopy techniques used for both ICSI and IMSI.


2022 ◽  
Vol 5 (1) ◽  
Author(s):  
Xavier Vendrell ◽  
Julia Ramírez-González ◽  
Zuo-Guang Ye ◽  
Anthony R. West

AbstractRelaxor ferroelectrics exhibit both static and dynamic local structural order which controls their frequency-dependent electrical properties. A combination of advanced scattering and microscopy techniques have been used recently to determine the local structure of relaxors. To complement these, here we show an approach to electrical property measurements which identifies local dipoles whose switching is co-operative, temperature-dependent and responsible for the observed dispersion in dielectric properties. Impedance measurements and equivalent circuit analysis of a canonical relaxor, Pb(Mg1/3Nb2/3)O3 single crystal, over the ranges 180–1050 K and 100 Hz–1 MHz, show that incorporation of a single constant phase element into the equivalent circuit used to fit experimental data is able to account fully for the dispersions that characterise the relaxor response, over this frequency range. This allows parametrisation of the relaxor behaviour, gives increased understanding of the relaxation mechanisms responsible and forms the basis for modifying and controlling relaxor characteristics.


2022 ◽  
Vol 66 (6) ◽  
pp. 434-441
Author(s):  
T. S. Trifonova ◽  
A. V. Moiseenko ◽  
M. V. Bourkaltseva ◽  
O. V. Shaburova ◽  
A. K. Shaytan ◽  
...  

Introduction. Giant phiKZ-like bacteriophages have a unique protein formation inside the capsid, an inner body (IB) with supercoiled DNA molecule wrapped around it. Standard cryo-electron microscopy (cryo-EM) approaches do not allow to distinguish this structure from the surrounding nucleic acid of the phage. We previously developed an analytical approach to visualize protein-DNA complexes on Escherichia coli bacterial cell slices using the chemical element phosphorus as a marker. In the study presented, we adapted this technique for much smaller objects, namely the capsids of phiKZ-like bacteriophages.Material and methods. Following electron microscopy techniques were used in the study: analytical (AEM) (electron energy loss spectroscopy, EELS), and cryo-EM (images of samples subjected to low and high dose of electron irradiation were compared).Results. We studied DNA packaging inside the capsids of giant bacteriophages phiEL from the Myoviridae family that infect Pseudomonas aeruginosa. Phosphorus distribution maps were obtained, showing an asymmetrical arrangement of DNA inside the capsid.Discussion. We developed and applied an IB imaging technique using a high angle dark-field detector (HAADF) and the STEM-EELS analytical approach. Phosphorus mapping by EELS and cryo-electron microscopy revealed a protein formation as IB within the phage phiEL capsid. The size of IB was estimated using theoretical calculations.Conclusion. The developed technique can be applied to study the distribution of phosphorus in other DNA- or RNA-containing viruses at relatively low concentrations of the element sought.


Author(s):  
V. S. M. Pereira ◽  
S. Wang ◽  
T. Morgan ◽  
H. Schut ◽  
J. Sietsma

AbstractIn the present work, an ODS 12 Cr steel was characterized using Electron Microscopy techniques, in an as-received condition and after annealing treatments between 773 K and 1573 K. Results show a complex microstructure, with the presence of fine Y–Ti–O nanoparticles dispersed in the matrix. After annealing at 1573 K, the average diameter of Y–Ti–O nanoparticles increases from ~ 4 to ~ 7 nm and partial recrystallization occurs. The trapping behavior of deuterium in the steel in its as-received state and annealed at 1573 K was investigated. Samples were exposed to low-energy deuterium plasma and analyzed with thermal desorption spectroscopy, after waiting times of 1 day and 25 days. The samples measured 1 day after exposure released a higher total amount of deuterium than the ones measured after 25 days. The effect of waiting time is explained by the release of deuterium, at 300 K, from sites with low activation energy for detrapping, Ed. In the as-received condition, part of the deuterium detrapped at 300 K was re-trapped by high-Ed sites. For the samples in the annealed condition, the redistribution of deuterium from low-Ed to high-Ed sites was not observed, but the total amount of deuterium released was higher.


2022 ◽  
Vol 13 ◽  
pp. 1-9
Author(s):  
Carl Drechsel ◽  
Philipp D’Astolfo ◽  
Jung-Ching Liu ◽  
Thilo Glatzel ◽  
Rémy Pawlak ◽  
...  

Topological superconductivity emerging in one- or two-dimensional hybrid materials is predicted as a key ingredient for quantum computing. However, not only the design of complex heterostructures is primordial for future applications but also the characterization of their electronic and structural properties at the atomic scale using the most advanced scanning probe microscopy techniques with functionalized tips. We report on the topographic signatures observed by scanning tunneling microscopy (STM) of carbon monoxide (CO) molecules, iron (Fe) atoms and sodium chloride (NaCl) islands deposited on superconducting Pb(111). For the CO adsorption a comparison with the Pb(110) substrate is demonstrated. We show a general propensity of these adsorbates to diffuse at low temperature under gentle scanning conditions. Our findings provide new insights into high-resolution probe microscopy imaging with terminated tips, decoupling atoms and molecules by NaCl islands or tip-induced lateral manipulation of iron atoms on top of the prototypical Pb(111) superconducting surface.


Antibiotics ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 11
Author(s):  
Alberto Vitali ◽  
Annarita Stringaro ◽  
Marisa Colone ◽  
Alexandra Muntiu ◽  
Letizia Angiolella

The increased prevalence and incidence of fungal infections, of which Candida albicans represents one of the most life-threatening organisms, is prompting the scientific community to develop novel antifungal molecules. Many essential oils components are attracting attention for their interesting antifungal activities. Given the chemical and physical characteristics of these compounds, the use of appropriate nanodelivery systems is becoming increasingly widespread. In this study, chitosan nanoparticles were prepared using an ionic gelation procedure and loaded with the phenolic monoterpene carvacrol. After a bioassay guided optimization, the best nanoparticle formulation was structurally characterized by means of different spectroscopic (UV, FTIR and DLS) and microscopy techniques (SEM) and described for their functional features (encapsulation efficiency, loading capacity and release kinetics). The antifungal activity of this formulation was assayed with different Candida spp., both in planktonic and biofilm forms. From these studies, it emerged that the carvacrol loaded nanoparticles were particularly active against planktonic forms and that the antibiofilm activity was highly dependent on the species tested, with the C. tropicalis and C. krusei strains resulting as the most susceptible.


Minerals ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 5
Author(s):  
Juan Sebastián Zuluaga-Morales ◽  
María Victoria Bolaños-Carmona ◽  
Carolina Cecilia Cifuentes-Jiménez ◽  
Pedro Álvarez-Lloret

In vitro simulation of natural caries is of great importance in dental research for the development of more effective clinical treatments. The pH-cycling (pHc) procedure consists of a dynamic caries process with alternating de-remineralisation periods. The current research aims to evaluate the effects of the pHc procedure on mineral dentine properties in comparison with sound dentine and natural residual caries. For this purpose, dentine slices from human third molars were submitted to cycling periods of 14 and 28 days. The chemical composition, morphological and microstructural properties of the dentine samples were examined by infrared and Raman spectroscopies, X-ray diffraction, and scanning electron microscopy techniques. In addition, the depth of the demineralisation front was evaluated by Masson’s trichrome (MT) staining. The results showed that the pHc procedure led to notable changes in the mineral composition and the crystalline characteristics with respect to sound dentine and some extent to natural caries. The MT results revealed that pHc 28 yields a deeper lesion than pHc 14, simulating potential progression of natural caries. The results of this study provide a better understanding of the mechanisms of demineralisation that could occur in an in vivo environment and provide a standardised substrate similar to natural residual caries.


Caryologia ◽  
2021 ◽  
Vol 74 (3) ◽  
pp. 91-97
Author(s):  
Ciler Kartal ◽  
Nuran Ekici ◽  
Almina Kargacıoğlu ◽  
Hazal Nurcan Ağırman

In this study gynoecium, megasporogenesis, megagametogenesis and female gametophyte of Gladiolus italicus Miller were examined cytologically and histologically by using light microscopy techniques. Ovules of G. italicus are of anatropous, bitegmic and crassinucellate type. Embryo sac development is of monosporic Polygonum type. Polar nuclei fuse before fertilization to form a secondary nucleus near the antipodals. The female gametophyte development of G. italicus was investigated for the first time with this study.


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