Search Results
Influence of nanobody binding on fluorescence emission, mobility, and organization of GFP-tagged proteins
2020, Schneider, Falk, Sych, Taras, Eggeling, Christian, Sezgin, Erdinc
Advanced fluorescence microscopy studies require specific and monovalent molecular labeling with bright and photostable fluorophores. This necessity led to the widespread use of fluorescently labeled nanobodies against commonly employed fluorescent proteins (FPs). However, very little is known how these nanobodies influence their target molecules. Here, we tested commercially available nanobodies and observed clear changes of the fluorescence properties, mobility and organization of green fluorescent protein (GFP) tagged proteins after labeling with the anti-GFP nanobody. Intriguingly, we did not observe any co-diffusion of fluorescently labeled nanobodies with the GFP-labeled proteins. Our results suggest significant binding of the nanobodies to a non-emissive, likely oligomerized, form of the FPs, promoting disassembly into monomeric form after binding. Our findings have significant implications on the application of nanobodies and GFP labeling for studying dynamic and quantitative protein organization in the plasma membrane of living cells using advanced imaging techniques.
Thermally induced evolution of the structure and optical properties of silicon nanowires
2020, Mussabek, Gauhar, Lysenko, Vladimir, Yermukhamed, Dana, Sivakov, Vladimir, Yu. Timoshenko, Victor
In the present paper, we report on the investigation of thermal annealing (TA) effect on structural and optical properties of crystalline silicon nanowires produced by metal-assisted chemical etching approach. In particular, the impact of TA on nanowire length, relative volume and size distribution of voids is described in terms of Lifshitz-Slyozov-Wagner theory considering the TA induced Oswald ripening in the SiNW arrays. It was also found that TA leads to a decrease of the SiNWs total reflection in the wide UV–VIS-IR spectral range. The reported effects can be used for tuning of crystalline SiNWs arrays in view of their further applications in photonics related fields. © 2020 The Authors
On the possibility of PhotoEmission Electron Microscopy for E. coli advanced studies
2020, Turishchev, S.Yu., Marchenko, D., Sivakov, V., Belikov, E.A., Chuvenkova, O.A., Parinova, E.V., Koyuda, D.A., Chumakov, R.G., Lebedev, A.M., Kulikova, T.V., Berezhnoy, A.A., Valiakhmedova, I.V., Praslova, N.V., Preobrazhenskaya, E.V., Antipov, S.S.
The novel approach was proposed for detailed high-resolution studies of morphology and physico-chemical properties concomitantly at one measurement spot of E. coli bacterial cells culture immobilized onto silicon wafer surface in UHV conditions applying PhotoEmission Electron Microscopy under Hg lamp irradiation. For the E. coli characterization scanning electron microscopy (electron beam) and X-ray photoelectron spectroscopy (X-ray tube radiation) were applied prior to PhotoEmission Electron Microscopy measurements. In spite of irradiation doses collected for the cell arrays we were successful in detection of high-resolution images even of single E. coli bacterium by PhotoEmission Electron Microscopy technique followed by detailed high-resolution morphology studies by scanning electron microscopy. These results revealed widespread stability of the E. coli membranes shape after the significant number of applied characterization techniques. © 2019 The Authors
Peculiarities of electronic structure and composition in ultrasound milled silicon nanowires
2020, Parinova, E.V., Pisliaruk, A.K., Schleusener, A., Koyuda, D.A., Chumakov, R.G., Lebedev, A.M., Ovsyannikov, R., Makarova, A., Smirnov, D., Sivakov, V., Turishchev, S.Yu.
The combined X-ray absorption and emission spectroscopy approach was applied for the detailed electronic structure and composition studies of silicon nanoparticles produced by the ultrasound milling of heavily and lowly doped Si nanowires formed by metal-assisted wet chemical etching. The ultrasoft X-ray emission spectroscopy and synchrotron based X-ray absorption near edges structure spectroscopy techniques were utilize to study the valence and conduction bands electronic structure together with developed surface phase composition qualitative analysis. Our achieved results based on the implemented surface sensitive techniques strongly suggest that nanoparticles under studies show a significant presence of the silicon suboxides depending on the pre-nature of initial Si wafers. The controlled variation of the Si nanoparticles surface composition and electronic structure, including band gap engineering, can open a new prospective for a wide range Si-based nanostructures application including the integration of such structures with organic or biological systems. © 2020