Atomic Force Microscopy Microscopy Research Articles

Development of Halloysite Nanohybrids-Based Films: Enhancing Mechanical and Hydrophilic Properties for Wound Healing.

Most of the therapeutic systems developed for managing chronic skin wounds lack adequate mechanical and hydration properties, primarily because they rely on a single component. This study addresses this issue by combining organic and inorganic materials to obtain hybrid films with enhanced mechanical behavior, adhesion, and fluid absorption properties. To that aim, chitosan/hydrolyzed collagen blends were mixed with halloysite/antimicrobial nanohybrids at 10% and 20% (w/w) using glycerin or glycerin/polyethylene glycol-1500 as plasticizers. The films were characterized through the use of Fourier-transform infrared (FTIR) spectroscopy, thermogravimetric analysis (TGA), and electron microscopy. The mechanical properties were evaluated macroscopically using tensile tests, and at a nanoscale through atomic force microscopy (AFM) and nanoindentation. Thermodynamic studies were conducted to assess their hydrophilic or hydrophobic character. Additionally, in vitro cytocompatibility tests were performed on human keratinocytes. Results from FTIR, TGA, AFM and electron microscopy confirmed the hybrid nature of the films. Both tensile tests and nanomechanical measurements postulated that the nanohybrids improved the films' toughness and adhesion and optimized the nanoindentation properties. All nanohybrid-loaded films were hydrophilic and non-cytotoxic, showcasing their potential for skin wound applications given their enhanced performance at the macro- and nanoscale.

Cytoarchitecture of Breast Cancer Cells under Diabetic Conditions: Role of Regulatory Kinases-Rho Kinase and Focal Adhesion Kinase.

Diabetes greatly reduces the survival rates in breast cancer patients due to chemoresistance and metastasis. Reorganization of the cytoskeleton is crucial to cell migration and metastasis. Regulatory cytoskeletal protein kinases such as the Rho kinase (ROCK) and focal adhesion kinase (FAK) play a key role in cell mobility and have been shown to be affected in cancer. It is hypothesized that diabetes/high-glucose conditions alter the cytoskeletal structure and, thus, the elasticity of breast cancer cells through the ROCK and FAK pathway, which can cause rapid metastasis of cancer. The aim of the study was to investigate the role of potential mediators that affect the morphology of cancer cells in diabetes, thus leading to aggressive cancer. Breast cancer cells (MDA-MB-231 and MCF-7) were treated with 5 mM glucose (low glucose) or 25 mM glucose (high glucose) in the presence of Rho kinase inhibitor (Y-27632, 10 mM) or FAK inhibitor (10 mM). Cell morphology and elasticity were monitored using atomic force microscopy (AFM), and actin staining was performed by fluorescence microscopy. For comparative study, normal mammary breast epithelial cells (MCF-10A) were used. It was observed that high-glucose treatments modified the cytoskeleton of the cells, as observed through AFM and fluorescence microscopy, and significantly reduced the elasticity of the cells. Blocking the ROCK or FAK pathway diminished the high-glucose effects. These changes were more evident in the breast cancer cells as compared to the normal cells. This study improves the knowledge on the cytoarchitecture properties of diabetic breast cancer cells and provides potential pathways that can be targeted to prevent such effects.

Aqueous Black Seed (Nigellasativa L.) Extract-Mediated Corrosion Inhibition in Mild Steel Exposed to 3.5% NaCl: Effect of Temperature, pH, Time, and In Situ Analysis Using Atomic Force Microscopy

Atomic force microscopy (AFM)-based analysis on the corrosion inhibition properties of aqueous black seed extract for mild steel exposed to 3.5% NaCl solution is reported here. At ~ 600 ppm inhibitor concentration, corrosion rate and inhibition efficiency are found to be ~ 0.18 ± 0.1 mpy and ~ 94%, respectively. The superior corrosion inhibition is due to the adsorption of the phytochemicals onto the metallic surface, as confirmed from spectroscopic analysis. Findings show the suitability of the black seed extract for practical applications at neutral pH and room temperature. AFM and magnetic force microscopy indicate lower surface roughness and nearly pristine topography for the inhibitor-protected specimens. In situ AFM analyses indicate delayed formation of surface irregularities for the inhibitor-protected specimens. The obtained results clearly show the superior corrosion inhibition efficiency of the aqueous black seed extract against mild steel under simulated sea water condition.

Nanoscale Structure of Lipid-Gemini Surfactant Mixed Monolayers Resolved with AFM and KPFM Microscopy.

Drug delivery vehicles composed of lipids and gemini surfactants (GS) are promising in gene therapy. Tuning the composition and properties of the delivery vehicle is important for the efficient load and delivery of DNA fragments (genes). In this paper, we studied novel gene delivery systems composed of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-3-phosphocholine (DPPC), and GS of the type N,N-bis(dimethylalkyl)-α,ω-alkanediammonium dibromide at different ratios. The nanoscale properties of the mixed DOPC-DPPC-GS monolayers on the surface of the gene delivery system were studied using atomic force microscopy (AFM) and Kelvin probe force microscopy (KPFM). We demonstrate that lipid-GS mixed monolayers result in the formation of nanoscale domains that vary in size, height, and electrical surface potential. We show that the presence of GS can impart significant changes to the domain topography and electrical surface potential compared to monolayers composed of lipids alone.

Molecular interaction of human serum albumin with gemini surfactant, 1,6 bis (N,N-hexadecyl dimethyl ammonium) bromide: Influence of micelles and pH

The interaction between human seram albumin (HSA) and a cationic gemini (dimeric) surfactant, 1,6 bis (N,N-hexadecyl dimethyl ammonium) bromide (16–6-16) has been investigated. Various analytical techniques, including UV–vis spectroscopy, fluorescence spectroscopy, lifetime fluorescence measurement, circular dichroism (CD), and atomic force microscope (AFM), were employed to investigate these interaction studies. The experiments were conducted in aqueous environments at three distinct pH levels (viz., below (4.0), at (4.7), and above (7.0) the isoelectric point (IP)), as well as both above and below the critical micellar concentration (CMC) of the surfactant (16–6-16). In these experimental conditions, HSA experiences conformational changes, leading to unfolding and denaturation as a result of its interaction with the gemini surfactant molecules (16–6-16). The extent of these conformational changes is found to be associated with both the degree of surfactant partial intercalation and the characteristics (size and charge) of the surfactant aggregates formed. The AFM microscopy studies provide insights into the topographical structural alterations occurring above, at, and below the CMC. Molecular docking analysis further validates the interaction between the protein (HSA) and the gemini surfactant (16–6-16) at a molecular level.

Shear-reversible clusters of HIV-1 in solution: stabilized by antibodies, dispersed by mucin.

The phenomenon of reversible clustering is expected to further nuance HIV immune stealth because virus surfaces can escape interaction with antibodies (Abs) by hiding temporarily within clusters. It is well known that mucin reduces HIV virulence, and the current perspective is that mucin aggregates HIV-1 to reduce infections. Our findings, however, suggest that mucin is dispersing HIV clusters. The study proposes a new paradigm for how HIV-1 may broadly evade Ab recognition with reversible clustering and why mucin effectively neutralizes HIV-1.

In situ mapping of biomineral skeletal proteins by molecular recognition imaging with antibody-functionalized AFM tips.

Spatial localizing of skeletal proteins in biogenic minerals remains a challenge in biomineralization research. To address this goal, we developed a novel in situ mapping technique based on molecular recognition measurements via atomic force microscopy (AFM), which requires three steps: (1) the development and purification of a polyclonal antibody elicited against the target protein, (2) its covalent coupling to a silicon nitride AFM tip ('functionalization'), and (3) scanning of an appropriately prepared biomineral surface. We applied this approach to a soluble shell protein - accripin11 - recently identified as a major component of the calcitic prisms of the fan mussel Pinna nobilis [1]. Multiple tests reveal that accripin11 is evenly distributed at the surface of the prisms and also present in the organic sheaths surrounding the calcitic prisms, indicating that this protein is both intra- and inter-crystalline. We observed that the adhesion force in transverse sections is about twice higher than in longitudinal sections, suggesting that accripin11 may exhibit preferred orientation in the biomineral. To our knowledge, this is the first time that a protein is localized by molecular recognition atomic force microscopy with antibody-functionalized tips in a biogenic mineral. The 'pros' and 'cons' of this methodology are discussed in comparison with more 'classical' approaches like immunogold. This technique, which leaves the surface to analyze clean, might prove useful for clinical tests on non-pathological (bone, teeth) or pathological (kidney stone) biomineralizations. Studies using implants with protein-doped calcium phosphate coating can also benefit from this technology. STATEMENT OF SIGNIFICANCE: Our paper deals with an unconventional technical approach for localizing proteins that are occluded in biominerals. This technique relies on the use of molecular recognition atomic force microscopy with antibody-functionalized tips. Although such approach has been employed in other system, this is the very first time that it is developed for biominerals. In comparison to more classical approaches (such as immunogold), AFM microscopy with antibody-functionalized tips allows higher magnification and keeps the scanned surface clean for other biophysical characterizations. Our method has a general scope as it can be applied in human health, for non-pathological (bone, teeth) and pathological (kidney stone) biomineralizations as well as for bone implants coated with protein-doped calcium phosphate.

Safety and Effectiveness of Conventional Commercial Products for Professional Tooth Bleaching: Comparative Ex Vivo Study Using AFM Microscopy and Nanoindentation

In modern times, patients are not only seeking oral health but also aiming for a flawless smile. Despite the progress made in aesthetic dentistry, there are instances where insufficient teeth whitening results contrast with the considerable cost of professional treatments. Patient discomfort and the potential for tooth damage further compound these challenges. In this study, we conducted a comparative split-mouth ex vivo investigation to evaluate the innovative BlancOne ULTRA+ (IDS SpA) in comparison with established professional products: Opalescence Extraboost (ULTRADENT), Zoom WhiteSpeed (Philips Research Eindhoven High Tech), and Pola Office (SDI Limited). Our initial focus was on the whitening effectiveness of each product, which was measured using a spectrophotometer. Subsequently, we assessed any structural changes in enamel post-treatment using an atomic force microscope (AFM) and a nanoindentation procedure. All tested bleaching agents demonstrated teeth-brightening effects. BlancOne ULTRA+, Zoom WhiteSpeed, and Pola Office caused minor alterations in the texture of the enamel surface within nanometric limits. However, Opalescence Extraboost exhibited notably more pronounced changes, indicating significant modifications in surface roughness and potential reductions in material hardness due to consequential shifts in mechanical properties. BlancOne ULTRA+ appears to offer the most favourable cost–benefit outcome.

In Situ Atomic Force Microscopy and X‐ray Computed Tomography Characterization of All‐Solid‐State Lithium Batteries: Both Local and Overall

All‐solid‐state lithium batteries (ASSLBs) are promising due to their high‐energy output and low‐risk profile, but their development has only just begun. Atomic force microscopy (AFM) and related techniques have had an impact on ASSLBs research by elucidating the interfacial, morphological, mechanical, electrical, and electrochemical properties of a wide range of electrodes and electrolytes. However, because a cross‐section cut is necessary to define the solid–solid interface, true in situ analysis is not practical. The first part of this review will assess recent advancements in the study of ASSLBs utilizing AFM and other scanning probe microscopy techniques. The interior solid–solid interfaces can be illuminated in situ using X‐ray computed tomography (X‐CT) and other nondestructive characterization techniques, whereas, in contrast, to deepen the subject, it is further examined how X‐CT vary from the use of other instruments for solid‐state battery characterization, compare the information that various methods may give, and assess how well they can accurately reflect real batteries. This review may serve as a reference and point researchers in the direction of future study on the solid–solid interface of ASSLBs.

Focus on lasers, imaging, microscopy, and nanoscience

Focus on lasers, imaging, microscopy, and nanoscience

3D Nanoprinting of All-Metal Nanoprobes for Electric AFM Modes.

3D nanoprinting via focused electron beam induced deposition (FEBID) is applied for fabrication of all-metal nanoprobes for atomic force microscopy (AFM)-based electrical operation modes. The 3D tip concept is based on a hollow-cone (HC) design, with all-metal material properties and apex radii in the sub-10 nm regime to allow for high-resolution imaging during morphological imaging, conductive AFM (CAFM) and electrostatic force microscopy (EFM). The study starts with design aspects to motivate the proposed HC architecture, followed by detailed fabrication characterization to identify and optimize FEBID process parameters. To arrive at desired material properties, e-beam assisted purification in low-pressure water atmospheres was applied at room temperature, which enabled the removal of carbon impurities from as-deposited structures. The microstructure of final HCs was analyzed via scanning transmission electron microscopy-high-angle annular dark field (STEM-HAADF), whereas electrical and mechanical properties were investigated in situ using micromanipulators. Finally, AFM/EFM/CAFM measurements were performed in comparison to non-functional, high-resolution tips and commercially available electric probes. In essence, we demonstrate that the proposed all-metal HCs provide the resolution capabilities of the former, with the electric conductivity of the latter onboard, combining both assets in one design.

Long and Covalently Conjugated Branched DNA Structures for in Situ Gelation in Vivo

Long and Covalently Conjugated Branched DNA Structures for in Situ Gelation in Vivo

Near-field infrared nanoscopic study of EUV- and e-beam-exposed hydrogen silsesquioxane photoresist

This article presents a technique of scattering-type scanning near-field optical microscopy (s-SNOM) based on scanning probe microscopy as a nanoscale-resolution chemical visualization technique of the structural changes in photoresist thin films. Chemical investigations were conducted in the nanometer regime by highly concentrated near-field infrared on the sharp apex of the metal-coated atomic force microscopy (AFM) tip. When s-SNOM was applied along with Fourier transform infrared spectroscopy to characterize the extreme UV- and electron-beam (e-beam)-exposed hydrogen silsesquioxane films, line and space patterns of half-pitch 100, 200, 300, and 500 nm could be successfully visualized prior to pattern development in the chemical solutions. The linewidth and line edge roughness values of the exposed domains obtained by s-SNOM were comparable to those extracted from the AFM and scanning electron microscopy images after development. The chemical analysis capabilities provided by s-SNOM provide new analytical opportunities that are not possible with traditional e-beam-based photoresist measurement, thus allowing information to be obtained without interference from non-photoreaction processes such as wet development.

Advancement and challenges in sample preparation for atomic force microscopy and infrared microscopy for wood-based materials.

Wood-based materials such as composites or laminates play an important role in today's furniture industry, especially in manufacturing high-quality kitchen and dining room furniture. One important aspect after fabrication is the investigation of these materials to derive quality metrics such as surface stain and scuff resistance. Current sample preparation methods are mostly straightforward and rely on cutting and grinding the materials under test, including sensitive wood substrates. To investigate cross-sections and different layer topologies, characterisation techniques such as atomic force microscopy (AFM) and infrared (IR) microscopy are of potential interest. However, a huge limitation is that current sample preparation methods lead to smeared coatings on the sample cross-sections and high surface roughness. Hence, these methods are not applicable for the sample preparation in measuring AFM and IR microscopy. Therefore, new preparation techniques need to be developed accordingly. This article presents a new approach towards coated wood-based sample preparation including embedding processes to use those samples for AFM and IR microscopy technologies. The proposed method has been evaluated by obtaining AFM, IR and microscopy measurements of more than four different wood-based samples such as (i) raw paper, (ii) impregnated paper, (iii) melamine-coated chipboards and (iv) medium-density fibreboards. The investigation results showed a significant improvement in sample preparation, as well as clear chemical and physical characterisation over whole sample construction, including coating layers, for wood-based materials.

Functional enhancement of hydroxypropyl cellulose‐based bionanocomposite films incorporating chitosan nanoparticles

SummaryThe aims of the present study were to investigate the effect of the incorporation of chitosan nanoparticles (ChiNPs) on the physicochemical properties and antifungal activity of hydroxypropyl cellulose (HPC) bionanocomposite films. The films of 4% HPC and ChiNP at various dosages (0%, 0.1%, 0.5%, 1%, 1.5%, 3% w/w of HPC) were prepared. Atomic force microscopy (AFM) verified the dispersion of spherical‐shaped ChiNP in a nanoscale size range (14.75–56.74 nm). Incorporation of nanofiller affected the functional properties by increasing the film–water contact angle to >53°, improving the optical properties, and reducing water vapour permeability by 5.73–12.54%. These results were influenced by surface roughness and microstructure characteristics of a bionanocomposite film, which were confirmed using AFM and scanning electron microscopy (SEM). Tensile assays of the films showed slight increases in tensile strength from 8.87 to 11.08 MPa and Young's modulus from 0.10 to 0.15 GPa with increasing proportions of ChiNP (1.5%). An effective concentration of ChiNP of 1.5% (w/w of HPC) was able to improve the performance of HPC and offered antifungal activity against Botrytis cinerea, as confirmed through mycelial growth inhibition assays and by membrane integrity changes. The HPC/ChiNP bionanocomposite films may have potential applications for active packaging, particularly for fresh fruit commodities.

Advances in Atomic Force Microscopy for the Electromechanical Characterization of Piezoelectric and Ferroelectric Nanomaterials

Given the social demand for self-powering wearable electronics, it is necessary to develop composite materials that exhibit both good flexibility and excellent piezoelectric performances. Intensive research on synthesis methods and devising characterization techniques for piezoelectric nanomaterials in various forms has been conducted. In particular, characterization techniques for piezoelectric nanomaterials require different approaches from those for conventional bulk materials. Atomic force microscopy (AFM)-based characterization techniques work based on the local physical interactions between the AFM tip and sample surfaces, making them an irreplaceable tool for studying the electromechanical properties of piezoelectric nanomaterials. Piezoresponse force microscopy (PFM), conductive AFM (C-AFM), and lateral force microscopy (LFM) are three representative AFM-based techniques used to characterize the piezoelectric and ferroelectric properties of nanomaterials. Coupled with the appearance of diverse novel nanomaterials such nanowires, free-standing nanorods, and electrospun nanofibers, AFM-based characterization techniques are becoming freer from artifacts and the need for quantitative measurements. PFM was initially developed to image the microstructures of piezoelectric materials, and well-calibrated techniques designed to realize quantitative measurements have been applied to nanomaterials. In contrast, C-AFM and LFM were initially used to measure the conductivity of diverse materials and the nanotribology of material surfaces. Over the last decade, they have proved their versatility and can now be used to evaluate the direct piezoelectric effect and the mechanical properties of piezoelectric nanomaterials. In these cases, systematic understanding with regard to the measurement principles is required for accurate measurements and analyses. In the present review article, we discuss earlier work in which AFM-based electromechanical characterization techniques were applied to nanomaterials to evaluate piezoelectric and ferroelectric properties. Also discussed is the importance of gaining a comprehensive understanding of the resulting signals.

Correlative AFM and Scanning Microlens Microscopy for Time-Efficient Multiscale Imaging.

With the rapid evolution of microelectronics and nanofabrication technologies, the feature sizes of large‐scale integrated circuits continue to move toward the nanoscale. There is a strong need to improve the quality and efficiency of integrated circuit inspection, but it remains a great challenge to provide both rapid imaging and circuit node‐level high‐resolution images simultaneously using a conventional microscope. This paper proposes a nondestructive, high‐throughput, multiscale correlation imaging method that combines atomic force microscopy (AFM) with microlens‐based scanning optical microscopy. In this method, a microlens is coupled to the end of the AFM cantilever and the sample‐facing side of the microlens contains a focused ion beam deposited tip which serves as the AFM scanning probe. The introduction of a microlens improves the imaging resolution of the AFM optical system, providing a 3–4× increase in optical imaging magnification while the scanning imaging throughput is improved ≈8×. The proposed method bridges the resolution gap between traditional optical imaging and AFM, achieves cross‐scale rapid imaging with micrometer to nanometer resolution, and improves the efficiency of AFM‐based large‐scale imaging and detection. Simultaneously, nanoscale‐level correlation between the acquired optical image and structure information is enabled by the method, providing a powerful tool for semiconductor device inspection.

Sub-micro scale cell segmentation using deep learning.

Automated cell segmentation is key for rapid and accurate investigation of cell responses. As instrumentation resolving power increases, clear delineation of newly revealed cellular features at the submicron through nanoscale becomes important. Reliance on the manual investigation of myriad small features retards investigation; however, use of deep learning methods has great potential to reveal cell features both at high accuracy and high speed, which may lead to new discoveries in the near term. In this study, semantic cell segmentation systems were investigated by implementing fully convolutional neural networks called U-nets for the segmentation of astrocytes cultured on poly-l-lysine-functionalized planar glass. The network hyperparameters were determined by changing the number of network layers, loss functions, and input image modalities. Atomic force microscopy (AFM) images were selected for investigation as these are inherently nanoscale and are also dimensional. AFM height, deflection, and friction images were used as inputs separately and together, and the segmentation performances were investigated on five-fold cross-validation data. Transfer learning methods, including VGG16, VGG19, and Xception, were used to improve cell segmentation performance. We find that AFM height images inherit more discriminative features than AFM deflection and AFM friction images for cell segmentation. When transfer-learning methods are applied, statistically significant segmentation performance improvements are observed. Segmentation performance was compared to classical image processing algorithms and other algorithms in use by considering both AFM and electron microscopy segmentation. An accuracy of 0.9849, Matthews correlation coefficient of 0.9218, and Dice's similarity coefficient of 0.9306 were obtained on the AFM test images. Performance evaluations show that the proposed system can be successfully used for AFM cell segmentation with high precision.

Tunable synthesis, characterization, and CMP performance of dendritic mesoporous silica nanospheres as functionalized abrasives

The type, morphology, structure, size and distribution, surface chemistry, and mechanical behavior of particle abrasives play a key role in advanced chemical mechanical polishing (CMP) applications. In this work, dendritic mesoporous silica (D-mSiO2) nanospheres with tunable structures were synthesized via a modified n-hexane/water biliquid system coupled with a shearing assisted interfacial coassembly method. The resulting samples were characterized in terms of transmission electron microscopy, scanning electron microscopy, thermogravimetric analysis, Fourier transform infrared spectroscopy, powder X-ray diffraction, and nitrogen adsorption-desorption measurements. The spherical D-mSiO2 products presented three-dimensional and center-radial structures with particle size of 30–140 nm, pore diameter of 3–9 nm, surface area of ~780 m2/g, and pore volume of ~1.5 cm3/g. The elastic property and compressive elastic modulus of D-mSiO2 nanospheres were investigated by atomic force microscopy (AFM) nanoindentation. Oxide-CMP performances of the series of D-mSiO2 nanospheres as functionalized abrasives were tracked via high-resolution AFM and interferometric microscopy. The subtle differences of surface characteristics were systematically compared in terms of roughness, topographical variation, and image surface area difference (SAD). Sub-100 nm D-mSiO2 nanospheres achieved superior CMP qualities with the decreased surface roughness (less than 0.2 nm Rq and 0.3 nm Rz, within 5.0 × 5.0 µm2), the reduced maximum peak height/minimum valley depth (less than 0.5 nm), as well as the low image SAD (less than 0.009%). Furthermore, the interfacial contact behavior, water-involved tribochemical wear, and material removal mechanism of D-mSiO2 nanoabrasives were also discussed. The investigations are expected to open up the possibility of utilizing D-mSiO2 nanospheres as novel abrasives for advanced CMP applications.

Fast and effective observations of the pore structure of tight sandstones at the same location by utilizing AFM and CF-SEM

Atomic force microscopy (AFM), which allows 3D construction, nondestructive, and has a high resolution has been used to describe the pore structure of unconventional reservoirs. Due to its scalability, convenience, and flexibility, scanning electron microscopy (SEM) is widely conducted to describe the surface morphology of unconventional reservoirs. However, the two technologies are rarely combined to achieve fast and effective observations of the pore structure of unconventional reservoirs at the same location of the same sample (SLSS). In this study, nanoindentation technology is introduced to create a marked district (MD). Afterward, fast and effective observations of the pore structure of tight sandstones at the SLSS can be realized by searching this identifiable district with AFM and cold-field scanning electron microscopy (CF-SEM). The results demonstrate that AFM and CF-SEM rapidly capture observations of the surface morphology of tight sandstones at the SLSS. The method used in this study accomplishes the goal of fast and effective scanning at the SLSS (only a few minutes), while exhibiting the high potential of combining AFM and CF-SEM for comprehensively investigating the pore structure of reservoirs ranging from nanoscale to microscale. The most unique component of the presented method is that it is especially for the in situ observation of variations in the surface properties (morphology, electric, mechanical, surface force, and roughness) and pore size of all heterogeneous materials subjected to various external stimuli because the applied experimental technologies (nanoindentation, AFM, and CF-SEM) are all exhibit nondestructive properties. • AFM and CF-SEM can be applied to observe the surface morphology at the same location. • Nanoindentation technology can be applied to create an identifiable district. • Fast and effective observations at the SLSS can be achieved. • In situ observations of heterogeneous materials treated by various external stimuli can be realized.