High-resolution Scanning Electron Micrographs Research Articles

Preparation of biological monolayers for producing high-resolution scanning electron micrographs.

Scanning electron microscopy (SEM) provides a technical platform for nanoscopic mapping of biological structures. Correct preparation of SEM samples can provide an unprecedented understanding of the nexus between cellular morphology and topography. This comparative study critically examines two coating methods for preparing biological samples for scanning electron microscopy, while also providing novel advice on how to prepare in vitro epithelial or endothelial samples for high-resolution scanning-electron microscopy (HR-SEM). Two obstacles often confront the biologist when investigating cellular structures grown under tissue culture conditions, namely., how to prepare and present the biological samples to the HR-SEM microscope without affecting topographical membrane and cellular structural alterations. Firstly, our use of the Millicell cellulose inserts on which to grow our cellular samples in preparation for HR-SEM is both novel and advantageous to comparing the permeability function of cells to their morphological function. Secondly, biological material is often non-conducting, thermally sensitive and fragile and, therefore, needs to be fixed correctly and coated with thin conducting metal to ensure high-resolution detail of samples. Immortalized mouse brain endothelial cells (bEnd5) was used as a basis for describing the preferences in the use of the protocol. We compare two biological sample coating modalities for the visualizing and analysis of texturized, topographical, membranous ultrastructures of brain endothelial cell (BEC) confluent monolayers, namely, carbon and gold:palladium (Au:Pd) sputter coating in preparation for HR-SEM. BEC monolayers sputter-coated with these two modalities produced three-dimensional micrographs which have distinctly different topographical detail from which the nanostructural cellular data can be examined. The two coating methods display differences in the amount of nanoscopic detail that could be resolved in the nanosized membrane cytoarchitecture of BEC monolayers. The micrographical data clearly showed that Au:Pd sputter-coated samples generate descript imagery, providing useful information for profiling membrane nanostructures compared to carbon-coated samples. The recommendations regarding the contrast in two modalities would provide the necessary guidance to biological microscopists in preparing tissue culture samples for HR-SEM.

Refutation of traumatic insemination in the Drosophila bipectinata species complex

Traumatic insemination (TI) is a rare reproductive behaviour characterized by the transfer of sperm to the female via puncture wounds inflicted across her body wall. Here, we challenge the claim made by Kamimura (Kamimura 2007 Biol. Lett. 3, 401–404. (doi:10.1098/rsbl.2007.0192)) that males of species of the Drosophila bipectinata complex use a pair of claw-like processes (claws) to traumatically inseminate females: the claws are purported to puncture the female body wall and genital tract, and to inject sperm through the wounds into the lumen of her genital tract, bypassing the vaginal opening. This supposed case of TI is widely cited and featured in prominent subject reviews. We examined high-resolution scanning electron micrographs of the claws and failed to discover any obvious ‘groove’ for sperm transport. We demonstrated that sperm occurred in the female reproductive tract as a single-integrated unit, inconsistent with the claim that sperm are injected via paired processes. Laser ablation of the sharp terminal ends of the claws failed to inhibit insemination. We showed that the aedeagus in the complex delivers sperm through the vaginal opening, as in other Drosophila. The results refute the claim of TI in the Drosophila bipectinata species complex.

Quantitative Morphology of Cerebral Thrombi Related to Intravital Contraction and Clinical Features of Ischemic Stroke.

The purpose was to assess quantitatively and qualitatively the composition and structure of cerebral thrombi and correlate them with the signs of intravital clot contraction (retraction), as well as with etiology, severity, duration, and outcomes of acute ischemic stroke. We quantified high-resolution scanning electron micrographs of 41 cerebral thrombi for their detailed cellular and noncellular composition and analyzed histological images for the overall structure with the emphasis on red blood cell compression, fibrin age, and the signs of inflammation. Cerebral thrombi were quite compact and had extremely low porosity. The prevailing cell type was polyhedral compressed erythrocytes (polyhedrocytes) in the core, and fibrin-platelet aggregates were concentrated at the periphery; both findings are indicative of intravital contraction of the thrombi. The content of polyhedrocytes directly correlated with the stroke severity. The prevalence of fibrin bundles was typical for more severe cases, while the content of fibrin sponge prevailed in cases with a more favorable course. The overall platelet content in cerebral thrombi was surprisingly small, while the higher content of platelet aggregates was a marker of stroke severity. Fibrillar types of fibrin prevailed in atherothrombogenic thrombi. Older fibrin prevailed in thrombi from the patients who received thrombolytics, and younger fibrin dominated in cardioembolic thrombi. Alternating layers of erythrocytes and fibrin mixed with platelets were common for thrombi from the patients with more favorable outcomes. Thrombi with a higher number of leukocytes were associated with fatal cases. Most cerebral thrombi undergo intravital clot contraction (retraction) that may be of underestimated clinical importance. Despite the high variability of the composition and structure of cerebral thrombi, the content of certain types of blood cells and fibrin structures combined with the morphological signs of intravital contraction correlate with the clinical course and outcomes of acute ischemic stroke.

Phytosynthesis of BiVO4 nanorods using Hyphaene thebaica for diverse biomedical applications

Biosynthesis of bismuth vanadate (BiVO4) nanorods was performed using dried fruit extracts of Hyphaene thebaica as a cost effective reducing and stabilizing agent. XRD, DRS, FTIR, zeta potential, Raman, HR-SEM, HR-TEM, EDS and SAED were used to study the main physical properties while the biological properties were established by performing diverse assays. The zeta potential is reported as − 5.21 mV. FTIR indicated Bi–O and V–O vibrations at 640 cm−1 and 700 cm−1/1120 cm−1. Characteristic Raman modes were observed at 166 cm−1, 325 cm−1 and 787 cm−1. High resolution scanning and transmission electron micrographs revealed a rod like morphology of the BiVO4. Bacillus subtilis, Klebsiella pneumonia, Fusarium solani indicated highest susceptibility to the different doses of BiVO4 nanorods. Significant protein kinase inhibition is reported for BiVO4 nanorods which suggests their potential anticancer properties. The nanorods revealed good DPPH free radical scavenging potential (48%) at 400 µg/mL while total antioxidant capacity of 59.8 µg AAE/mg was revealed at 400 µg/mL. No antiviral activity is reported on sabin like polio virus. Overall excellent biological properties are reported. We have shown that green synthesis can replace well established processes for synthesizing BiVO4 nanorods.

Impact of Plasmon-Induced Atoms Migration in Harmonic Generation

Under illumination of a Ti:sapphire femtosecond oscillator, amplification of third harmonic generation by subwavelength plasmonic apertures is observed. However, the harmonic yield efficiency decays rapidly over time. In this work we investigate the physical phenomena behind the temporal attenuation of the harmonic signal. From high-resolution scanning electron micrographs and two-dimensional energy dispersive X-ray maps, we conclude that the attenuation of the third harmonic is attributed to trapping of a low-density carbon layer inside the plasmonic apertures. Furthermore, we show that the profile of the carbon deposit follows the enhanced electric near-field distribution, which indicates that the carbon atoms are transported to the field hotspot by the plasmonically enhanced optical tweezer effect. From the measurement of linear transmission spectra, we find that the dielectric constant inside the nanoholes is increased by the carbon deposit. However, numerical simulations demonstrate that the increase...

Delayed condensation and frost formation on superhydrophobic carbon soot coatings by controlling the presence of hydrophilic active sites

Condensation frosting is an undesired natural phenomenon that could be impeded efficiently using appropriate wettability and morphologically patterned surfaces. The icephobic properties of carbon soot and the fabrication scalability of its synthesis method are a good foundation for anti-frosting applications; however, the fundamentals of frost growth and spreading on sooted surfaces have not been examined yet. In this study, we investigate the anti-frosting performance of three groups of superhydrophobic soot coatings by means of 16 MHz quartz crystal microbalances (QCMs). The analysis of the real-time sensor signal of each soot coated QCM pattern shows that frost formation and its propagation velocity depend on the quantity of oxygen functionalities and structural defects in the material. In turn, the reduction of both parameters shifts the onset of frost growth to temperatures below −20 °C, whereas the interdroplet ice bridging is slowed by a factor of four. Moreover, high-resolution scanning electron micrographs of the samples imply delamination upon defrosting of the soot with spherical-like morphology via polar interactions driven mechanism. These results reveal an opportunity for control of frost incipiency on sooted surfaces by adjusting the synthesis conditions and depositing soot coatings with as low as possible content of hydrophilic active sites.

Pollen morphology of extant Winteraceae: a study allowing SEM-based affiliation of its fossil representatives

AbstractWhen applying high-resolution microscopy, the pollen morphology of extant taxa can be used to classify fossil pollen, that is, to address the latter in the established systematic-phylogenetic framework. Here we investigate tetrads and pollen features of 20 different Winteraceae species, most of them belonging to the early-diverging generic lineagesTasmannia,DrimysandPseudowintera. The tetrads and pollen are grouped into eleven pollen types based on diagnostic features observed by both light and scanning electron microscopy. The high-resolution scanning electron micrographs of recent material allow detailed comparison with fossil material, resulting in a more accurate affiliation of fossil tetrads/pollen to extant lineages. As a case study, early Miocene Winteraceae tetrads from South Africa are re-examined and formally described. The systematic placement of the African fossils is discussed in light of the pollen types presented here.

Self-optimization of plasmonic nanoantennas in strong femtosecond fields

Plasmonic dimer nanoantennas can significantly boost the electric field strength in the gap region, allowing for a modification of the feed gap geometry by femtosecond laser illumination. Using resonant bowtie antennas to enhance the electric field of a low-fluence femtosecond oscillator, here we experimentally demonstrate highly localized reshaping of the antennas, resulting in a self-optimization of the antenna shape. From high-resolution scanning electron micrographs and two-dimensional energy dispersive x-ray maps, we analyze the near-field enhanced subwavelength ablation at the nanotips and the resulting deposition of ablated materials in the feed gap. The dominant ablation mechanism is attributed to the nonthermal transient unbonding of atoms and electrostatic acceleration of ions. This process is driven by surface plasmon enhanced electron emission, with subsequent acceleration in the vacuum. This ablation is impeded in the presence of an ambient gas. A maximum of sixfold enhancement of the third-harmonic yield is observed during the reshaping process.

The effect of polyacrylic acid and reaction conditions on nanocluster formation of precipitated calcium carbonate on microcellulose

The precipitation of micro- and nanoparticles of calcium carbonate onto lignocellulosic microfibers was investigated at different microfiber concentrations with and without polyacrylic acid (PAA), i.e. a polymer commonly used to form polymer-induced liquid precursors of CaCO3. Concentrations of PAA, Ca(OH)2, CO2 and microfiber were varied in order to study the impact of reaction conditions on PCC formation in a batch reactor operated at ambient temperature. High resolution scanning electron micrographs of the samples show that both microfiber concentration and PAA dosage affected the nucleation and crystal growth of PCC filler on cellulosic fiber. Interestingly, at higher microfiber concentrations, larger amount of nano-sized spherical crystals were formed on the microfibers. A higher dosage of PAA, on the other hand, resulted in less nucleation on the microfiber, suggesting a preferential bulk nucleation mechanism. A higher concentration of PAA during the precipitation also led to the formation and stabilization of amorphous CaCO3, which was supported by SEM images and XRD analysis (lack of characteristic crystal structure).

Activated Charcoal Supported Cadmium Doped TiO<sub>2</sub> for Photocatalytic and Antibacterial Applications

The photocatalytic activity was studied under UV light using AC-Cd/TiO2 prepared via precipitation method and characterized by powder X-ray diffraction (XRD) , high resolution scanning electron micrographs (HR-SEM) with energy dispersive X-ray analysis (EDX), photoluminescence (PL) and Fourier transform Raman analysis (FT-RAMAN). The enhanced photo catalytic activity of the AC-Cd/TiO2 is demonstrated through photodegradation of methylene blue under UV light radiation at 365 nm. The mechanism of photocatalytic effect of AC-Cd/TiO2 nanocomposite material has been discussed Further its antibacterial activity against two gram positive and two gram negative bacterial strain is studied.

Activated Charcoal Supported Cadmium Doped TiO<sub>2</sub> for Photocatalytic and Antibacterial Applications

The photocatalytic activity was studied under UV light using AC-Cd/TiO2 prepared via precipitation method and characterized by powder X-ray diffraction (XRD) , high resolution scanning electron micrographs (HR-SEM) with energy dispersive X-ray analysis (EDX), photoluminescence (PL) and Fourier transform Raman analysis (FT-RAMAN). The enhanced photo catalytic activity of the AC-Cd/TiO2 is demonstrated through photodegradation of methylene blue under UV light radiation at 365 nm. The mechanism of photocatalytic effect of AC-Cd/TiO2 nanocomposite material has been discussed Further its antibacterial activity against two gram positive and two gram negative bacterial strain is studied.

A fundamental understanding of the dependence of the laser-induced breakdown spectroscopy (LIBS) signal strength on the complex focusing dynamics of femtosecond laser pulses on either side of the focus.

We correlate the focusing dynamics of 50 femtosecond (fs) laser radiation as it interacts with a silicon sample to laser-induced breakdown spectroscopy (LIBS) signal strength. Presented are concentric ring-shaped variations in the electric field in the prefocus region due to lens aberrations and nonsymmetry between the prefocus and post-focus beam profile as a result of continuum generation, occurring around the focus. Experimental results show different signal trends for both atmospheric and vacuum conditions, attributed to the existence of a continuum for the former. Lens aberrations effects on the LIBS signal strength are investigated using a plano-convex spherical lens and an aspherized achromatic lens. High-resolution scanning electron micrographs of the silicon surface after ablation, along with theoretical simulations, reveal the electric field patterns near the focus. The research results contribute to fundamental understanding of the basic physics of ultrashort, femtosecond laser radiation interacting with materials.

Particulate sol–gel synthesis and optical and electrical properties of CeO2/TiO2 nanocomposite

CeO2/TiO2 nanocomposite was synthesized by particulate sol–gel method. The X-ray diffractogram shows the presence of cubic CeO2 and anatase TiO2 in the composite. The high resolution scanning electron micrographs reveal the nanoparticulate nature of the prepared composite. The composite absorbs UV light and exhibits near-band gap emission corresponding to TiO2 and deep level emission due to crystal defects. The Nyquist plot displays two semicircular arcs indicating the material heterogeneity. The physicochemical characteristics of the synthesized nanocomposite are in favour of its application as an ingredient of sunscreen formulations; under UV light the photocatalytic activity of CeO2/TiO2 composite, tested through the degradation of rhodamine B, is very much less than that by pristine anatase TiO2. Reduced adsorption of moisture by the nanocomposite is a possible reason for the observed very low photocatalytic activity.

Outer shell, inner blocklets, and granule architecture of potato starch

Potato starch was heated in distilled water and acetic acid solution (pH=1.5), respectively, then washed with anhydrous ethanol and dried. After cooked in water with certain program, high-resolution scanning electron micrographs (SEM) of broken granules showed isolated outer shells. After treatment with acetic acid, SEM images of dissilient granules exhibited outer shells with randomly assembled spherical inner blocklets. A refined blocklet model was proposed for potato starch granule, the smaller blocklets were compacted densely to form the stiffer outer shell, and the larger blocklets were packed loosely to form the inner shells.

Nanostructured CexZn1−xO thin films: Influence of Ce doping on the structural, optical and electrical properties

Thin films of CexZn1−xO thin films were deposited on glass substrates at 400°C by nebulizer spray pyrolysis technique. Ce doping concentration (x) was varied from 0 to 10%, in steps of 2.5%. X-ray diffraction reveals that all the films have polycrystalline nature with hexagonal crystal structure and high preferential orientation along (002) plane. Optical parameters such as; transmittance, band gap energy, refractive index (n), extinction coefficient (k), complex dielectric constants (εr, εi) and optical conductivity (σr, σi) have been determined and discussed with respect to Ce concentration. All the films exhibit transmittance above 80% in the wavelength range from 330 to 2500nm. Optical transmission measurements indicate the decrease of direct band gap energy from 3.26 to 3.12eV with the increase of Ce concentration. Photoluminescence spectra show strong near band edge emission centered ∼398nm and green emission centered ∼528nm with excitation wavelength ∼350nm. High resolution scanning electron micrographs indicate the formation of vertical nano-rod like structures on the film surface with average diameter ∼41nm. Electrical properties of the Ce doped ZnO film have been studied using ac impedance spectroscopy in the frequency range from 100Hz–1MHz at different temperatures.

Application of response surface method on machining of Al–SiC nano-composites

The newly fabricated metal matrix nano-composite (MMNC) of Al 7075 reinforced with 1.5wt% SiC nano-particles was prepared by a novel ultrasonic cavitation method. The high resolution scanning electron micrograph (SEM) and field emission scanning electron micrograph (FESEM) shows uniform distribution and good dispersion of the SiC nanoparticles within the aluminum metal matrix. Electrical discharge machining (EDM) was employed to machine MMNC with copper electrode by adopting face centered central composite design of response surface methodology. Analysis of variance was applied to investigate the influence of process parameters and their interactions. Further a mathematical model has been formulated in order to estimate the machining characteristics. It has been observed that pulse current was found to be the most important factor affecting all the three output parameters such as material removal rate (MRR), electrode wear rate (EWR) and surface roughness (SR). The optimum parameter of combination setting has been identified for the MMNC are voltage 50.00V, pulse current 8.00A, Pulse on time 8.00μs and pulse off time 9.00μs. Finally the parameters were optimized for maximizing MRR, minimizing EWR and SR using desirability function approach.

Immobilized Nanorod Assemblies: Fabrication and Understanding of Large Area Surface-Enhanced Raman Spectroscopy Substrates

We describe the fabrication of optimized plasmonic substrates in the form of immobilized nanorod assemblies (INRA) for surface-enhanced Raman spectroscopy (SERS). Included are high-resolution scanning electron micrograph (SEM) images of the surface structures, along with a mechanistic description of their growth. It is shown that, by varying the size of support microspheres, the surface plasmon resonance is tuned between 330 and 1840 nm. Notably, there are predicted optimal microsphere sizes for each of the commonly used SERS laser wavelengths of 532, 633, 785, and 1064 nm.

Modeling and numerical simulations of growth and morphologies of three dimensional aggregated silver films

We report on a model for the deposition of three dimensional, aggregated nanocrystalline silver films, and an efficient numerical simulation method developed for visualizing such structures. We compare our results to a model system comprising chemically deposited silver films with morphologies ranging from dilute, uniform distributions of nanoparticles to highly porous aggregated networks. Disordered silver films grown in solution on silica substrates are characterized using digital image analysis of high resolution scanning electron micrographs. While the latter technique provides little volume information, plane-projected (two dimensional) island structure and surface coverage may be reliably determined. Three parameters governing film growth are evaluated using these data and used as inputs for the deposition model, greatly reducing computing requirements while still providing direct access to the complete (bulk) structure of the films throughout the growth process. We also show how valuable three dimensional characteristics of the deposited materials can be extracted using the simulated structures.

EDM of cast Al/SiC metal matrix nanocomposites by applying response surface method

The newly engineered metal matrix nanocomposite (MMNC) of Al 7075 reinforced with 0.5 wt% SiC nanoparticles was prepared by ultrasonic cavitation method. The high resolution scanning electron micrograph shows uniform distribution and good dispersion of the SiC nanoparticles within the aluminum metal matrix. Electrical discharge machining was employed to machine MMNC with copper electrode by adopting face-centered central composite design of response surface methodology. Analysis of variance was applied to investigate the influence of process parameters and their interactions. Further, a mathematical model has been formulated in order to estimate the machining characteristics.

Photocatalytic and bactericidal activities of hydrothermally synthesized nanocrystalline Cd-doped ZnO

Nanocrystalline Cd-doped ZnO has been obtained by hydrothermal synthesis and characterized by powder X-ray diffraction, energy dispersive X-ray spectrum, high resolution scanning electron micrographs and UV–visible diffuse reflectance, photoluminescence and electrochemical impedance spectra. Cd-doping by hydrothermal method decreases the grain size and destroys the microstructure. Although doping does not modify the band gap it suppresses the deep level emission, decreases the charge-transfer resistance and increases the capacitance. Cd-doping enhances the photocatalytic and bactericidal activities.