Journal of microscopy最新文献

Interferometric scattering (iSCAT) microscopy enables the label-free observation of biomolecules. Consequently, single-particle imaging and tracking with the iSCAT-based method known as mass photometry (MP) is a growing area of study. However, establishing reliable cover glass passivation and functionalisation methods is crucial to reduce nonspecific binding and prepare surfaces for in vitro single-molecule binding experiments. Existing protocols for fluorescence microscopy can contain strongly scattering or mobile components, which make them impractical for MP-based microscopy. In this study, we characterise several different surface coatings using MP. We present approaches for cover glass passivation using 3-aminopropyltriethoxysilane (APTES) and polyethylene glycol (PEG, 2k) along with functionalisation via a maleimide-thiol linker. These coatings are compatible with water or salt buffers, and show low background scattering; thus, we are able to measure proteins as small as 60 kDa. In this technical note, we offer a surface preparation suitable for in vitro experiments with MP.

Engineered cementitious composites (ECC) are a class of high-performing fibre-reinforced cementitious materials recognised for their increased ductility and durability compared to conventional cement-based materials, owing to their autogenously controlled tight crack widths, even when subjected to high strains. To reduce ECC's environmental impact, this research examines the use of a low-clinker binder − limestone-calcined clay cement (LC3) − as an alternative to portland cement (PC), along with fly ash to further reduce the clinker proportion and the embodied CO₂ of the formulations. In conventional concrete, LC3 hydrates to a denser microstructure resulting from the synergistic reaction between limestone and calcined clay. At the lower water contents typical of ECC and with the presence of fly ash, the influence of the binder composition on the microstructure is difficult to anticipate.

To examine the influence of these compositional variables on microstructure, permeability and durability, the sulphate resistance of LC3-based ECC is explored. Specifically, the ECC-LC3 blends are designed with high clinker replacement rate of 75% by mass of binder and contain either conventional fly ash or reclaimed fly ash at 50% by mass of binder. Expansion of ECC-LC3 samples subjected to standard sodium sulphate test conditions was measured up to 12 months and the depth of penetration of sulphates into the ECC-LC3 of varying compositions was quantified using micro-X-Ray Fluorescence (microXRF) imaging and modelling. The expansion results show that the ECC-LC3 formulations performed better than the PC samples and can provide adequate resistance to external sulphate attack, even when reclaimed fly ashes are used in place of the conventional ash. In addition, the shallow penetration of sulphate into these cementitious composites demonstrates the low diffusion coefficients values that were determined using the quantitative data from MicroXRF imaging.

We present a comparison of common electron microscopy sample preparation methods for studying crystallisation processes from solution using both scanning and transmission electron microscopy (SEM and TEM). We focus on two widely studied inorganic systems: calcium sulphate, gypsum (CaSO₄·2H₂O) and calcium carbonate (CaCO₃). We find significant differences in crystallisation kinetics and polymorph selection between the different sample preparation methods, which indicate that drying and chemical quenching can induce severe artefacts that are capable of masking the true native state of the crystallising solution. Overall, these results highlight the importance of cryogenic (cryo)-quenching crystallising solutions and the use of full cryo-TEM as the most reliable method for studying the early stages of crystallisation.

The performance of cementitious composites reinforced with fibres or/and bars depends on the bond strength between inclusion and cementitious matrix. The nature of formation of fibre-matrix bond is crucial for enhancing the reliability and utilisation of reinforced composites. The research provides a review on the recently published result about the changes in the microstructure of cement matrix surrounding steel fibres with different surface roughness, using a scanning electron microscope (SEM) coupled with k-means clustering algorithm for image segmentation. The debonding pattern of the fibre-matrix bond after the tensile loading cycles was discussed by observing the amount of adhered cement paste to the pulled out fibre surface with SEM. Therefore, analysis of SEM images enabled to explain the connection between the micro-scale properties of cement paste and fibre after application of cyclic loading.

Fibre bundle (FB)-based endoscopes are indispensable in biology and medical science due to their minimally invasive nature. However, resolution and contrast for fluorescence imaging are limited due to characteristic features of the FBs, such as low numerical aperture (NA) and individual fibre core sizes. In this study, we improved the resolution and contrast of sample fluorescence images acquired using in-house fabricated high-NA FBs by utilising generative adversarial networks (GANs). In order to train our deep learning model, we built an FB-based multifocal structured illumination microscope (MSIM) based on a digital micromirror device (DMD) which improves the resolution and the contrast substantially compared to basic FB-based fluorescence microscopes. After network training, the GAN model, employing image-to-image translation techniques, effectively transformed wide-field images into high-resolution MSIM images without the need for any additional optical hardware. The results demonstrated that GAN-generated outputs significantly enhanced both contrast and resolution compared to the original wide-field images. These findings highlight the potential of GAN-based models trained using MSIM data to enhance resolution and contrast in wide-field imaging for fibre bundle-based fluorescence microscopy.

Lay Description: Fibre bundle (FB) endoscopes are essential in biology and medicine but suffer from limited resolution and contrast for fluorescence imaging. Here we improved these limitations using high-NA FBs and generative adversarial networks (GANs). We trained a GAN model with data from an FB-based multifocal structured illumination microscope (MSIM) to enhance resolution and contrast without additional optical hardware. Results showed significant enhancement in contrast and resolution, showcasing the potential of GAN-based models for fibre bundle-based fluorescence microscopy.

Both copper and lithium act as strong fluxes and lower the temperature of the clinker melt formation. Sulphur promotes the stabilisation of more hydraulically active modification of alite M₁. It is expected that this combination could produce an alite clinker at significantly lower temperatures with high quality technological parameters. In this paper, the effect of combined oxides of copper, lithium and sulphur addition on the phase composition and clinker structure of Portland cement was investigated. The reference raw meal was prepared from common cement raw materials. Each of the mentioned oxides was added to the reference raw meal in two different concentrations, and 8 combinations were prepared. Chemically pure compounds (NH₄)₂SO₄, CuO and Li₂CO₃ were used as a source of these oxides. The raw meals were burned to equilibrium at 1450°C. Their phase composition was determined by X-ray diffraction analysis, the microstructure was monitored by optical microscopy, and the microchemistry of the clinker phases was observed by electron microscopy with EDS analysis. It was found that in samples with high lithium or copper content, there is an increase in belite and free lime at the expense of alite. The combination of Cu + Li has the most negative effect, followed by Li alone and Cu alone. The higher SO₃ content slightly offsets this negative effect.

Global efforts to minimise carbon dioxide emissions are also leading to attempts to use calcined clays (CC) as a partial substitute for cement in concrete. While the hydration mechanism of such CC blended cements is now well understood, the range of effective admixtures like polycarboxylate ethers (PCE) is limited. There are PCE types that promise relatively high effectiveness, but the mechanisms of action are not yet sufficiently understood. For a detailed understanding of the adsorption of such PCEs, spatially resolved studies of the binder were performed using a combination of fluorescence and scanning electron microscopy. In a comparison of two superplasticisers, the investigations have shown different sites of preferred adsorption in a CC blended system and the results can be correlated with flow tests and setting behaviour. The investigations have shown that a certain PCE type has a higher adsorption on CC and other components of a blended system in comparison to other types.