Polyphenols as Morphogenetic Agents for the Controlled Synthesis of Mesoporous Silica Nanoparticles

Non-surfactant-induced synthesis of mesoporous silica nanoparticles (MSNPs) is gaining increasing interest because of their low toxicity and simple purification compared to conventional surfactant-based methods. Tannic acid (TA), considered as a glucose-derived polyphenol, was first employed a few years ago and has attracted great research interest. Despite recent progress, the mechanisms resulting in the porous structure remain to be elucidated. In this work, we have employed TA and four structurally related polyphenols (gallic acid, ethyl gallate, eudesmic acid, and quercetin) to elucidate the effect of the chemical structure and properties of polyphenols on their templating ability. Our results unravel the mechanism of MSNP formation templated by TA, which form a supramolecular framework as the skeleton for the silica species to attach. The structure of the supramolecular network results in irregular pores. Additionally, the pK(a) value of the templates may be accounted for the particle size. Small-angle X-ray scattering was used to provide precise information on the morphology, especially the porosity of the resulting MSNPs in addition to electron microscopy, dynamic light scattering, nitrogen adsorption and desorption Brunauer-Emmett-Teller method, and thermogravimetric analysis.

Low-energy Electron Holography Microscope for Imaging of Single Molecules

Sven Alexander Szilagyi

Structure determination techniques are an essential tool to investigate and understand molecules, especially in biology, where functionality crucially depends on structure. The dominant high-resolution methods, such as transmission electron microscopy and X-ray diffraction, rely on ensemble averaging, making them unsuitable for evidencing structural variations of flexible molecules. Common single-molecule imaging techniques like scanning probe microscopy achieve high resolution but lack access to 3D objects. A possible way to overcome these limitations is offered by low-energy electron holography (LEEH). So far, LEEH achieved molecular imaging at resolutions in the range of 7-8 Å on the single-molecule level. A resolution in the range of a few Å, which would give access to atomic details of molecular structures, is in theory attainable. Obtaining a resolution near the theoretical limit requires a fully optimized LEEH setup and workflow, for which in particular sample and emitter preparation are crucial. Here, I present a novel LEEH microscope with a unique experimental workflow, which is capable of resolving molecular substructures with an unprecedented resolution for LEEH experiments. The newly constructed LEEH microscope takes advantage of an efficient vibration isolation system and is remote-controlled to minimize external perturbations. The setup is capable of low-temperature (50 K) measurements, which enhances the stability of the system. A reliable way for preparing highly coherent electron emitters constituted by sharp tungsten tips is presented along with different methods for improving their performance via functionalization and UHV treatments. I show an optimized and reliable substrate preparation protocol resulting in ultra-clean free-standing single-layer graphene (SLG) samples. Native electrospray ion beam deposition (ES-IBD) allows for the transfer of non-volatile molecules from solution into gas phase and onto the substrate. By employing a quadrupole mass selection, highly pure molecular ion beams are obtained, which are soft-landed onto the SLG substrate in a controlled and reproducible manner while retaining an intact structure. This versatile preparation method allows us to study a great variety of molecules by LEEH. Data on proteins with masses ranging from 12-820 kDa reveal molecular shapes consistent with model structures. Locally, structural details as small as 5 Å were identified, thus we are approaching our goal of a resolution in the range of a few Å. The application of our methodology to flexible proteins, exemplified by antibodies, allowed for the imaging of their conformational variability and for determining the influence of the sample preparation on their structure. This gave us the possibility to directly image gas-phase related conformations. Data of compact molecular systems with masses below 3 kDa reveal that our LEEH microscope is able to investigate small molecules, such as porphyrines, at high imaging contrast. The presented results indicate that an optimized LEEH setup and workflow in combination with ES-IBD is a versatile microscopy method, which is filling the gap between high-resolution and single-molecule techniques by elucidating structural details of various molecular species.

EPFL2022

Growth study of indium-catalyzed silicon nanowires by plasma enhanced chemical vapor deposition

Sonia Conesa Boj, Anna Fontcuberta i Morral, Père Roca i Cabarrocas, Libo Yu

Indium was used as a catalyst for the synthesis of silicon nanowires in a plasma enhanced chemical vapor deposition reactor. In order to foster the catalytic activity of indium, the indium droplets had to be exposed to a hydrogen plasma prior to nanowire growth in a silane plasma. The structure of the nanowires was investigated as a function of the growth conditions by electron microscopy and Raman spectroscopy. The nanowires were found to crystallize along the < 111 >, < 112 > or < 001 > growth direction. When growing on the < 112 > and < 111 > directions, they revealed a similar crystal quality and the presence of a high density of twins along the {111} planes. The high density and periodicity of these twins lead to the formation of hexagonal domains inside the cubic structure. The corresponding Raman signature was found to be a peak at 495 cm(-1), in agreement with previous studies. Finally, electron energy loss spectroscopy indicates an occasional migration of indium during growth.

2010

Identification of a Dps contamination in Mitomycin-C–induced expression of Colicin Ia

Henning Paul-Julius Stahlberg

Colicins are bacterial toxins targeting Gram-negative bacteria, including E. coli and related Enterobacteriaceae strains. Some colicins form ion-gated pores in the inner membrane of attacked bacteria that are lethal to their target. Colicin Ia was the first pore-forming E. coli toxin, for which a high-resolution structure of the monomeric full-length protein was determined. It is so far also the only colicin, for which a low-resolution structure of its membrane-inserted pore was reported by negative-stain electron microscopy. Resolving this structure at the atomic level would allow an understanding of the mechanism of toxin pore formation. Here, we report an observation that we made during an attempt to determine the Colicin Ia pore structure at atomic resolution. Colicin Ia was natively expressed by mitomycin-C induction under a native SOS promotor and purified following published protocols. The visual appearance in the electron microscope of negatively stained preparations and the lattice parameters of 2D crystals obtained from the material were highly similar to those reported earlier resulting from the same purification protocol. However, a higher-resolution structural analysis revealed that the protein is Dps (DNA-binding protein from starved cells), a dodecameric E. coli protein. This finding suggests that the previously reported low-resolution structure of a “Colicin Ia oligomeric pore” actually shows Dps.

2021

Low-energy Electron Holography Microscope for Imaging of Single Molecules

Sven Alexander Szilagyi

EPFL2022