Ordinary Microscope Research Articles

Morphological Characterization of Sb-Doped SnO2 Thin Films Developed by Spray Pyrolysis

In the present work, we have successfully deposited Sb antimony doped tin dioxide (SnO2) thin films with different concentrations (0 at%, 1 at%, 3 at% and 5 at%) from a stannous (II) chloride dihydrate solution (SnCl2.2H2O), by the pyrolysis chemical spray deposition technique on ordinary microscope glass substrates preheated to a fixed temperature of 350 °C. After deposition, the films were annealed at temperatures 400 °C for 4h. The aim of this work is on the one hand to study the morphological properties of pure and doped tin dioxide thin films and to determine the different morphological parameters such as: the roughness Ra, Rq and on the other hand to study the effect of Sb doping on the morphological properties of SnO2 thin films. To achieve this purpose, the thickness of all the films was estimated by profilometer with standard scan type. The morphology of the films was examined by atomic force microscope (AFM) in contact mode at room temperature to visualize the surface of our samples (the structure, size and morphology of crystallites ... etc). The 2D and 3D AFM images confirm the formation of nanostructures on the surface, with shapes and dimensions influenced by the amount of antimony doping. All the samples show a polycrystalline morphology, and the grain size progressively decreases with the increase of the Sb concentration (1% and 3%). On the other hand, for high antimony doping (5% Sb) the grains are larger than those observed at 3% doping. The root mean square (RMS) surface roughness values for samples with Sb concentrations (5%) were found to be 8.334 nm.

Single-Cell Sensitive Colorimetric pH Detection Based on Microscope Ratiometric Grayscale.

Rapid and accurate identification of the intracellular pH is critical in the field of biomedicine. In this work, we effectively identified and quantified the intracellular pH and its distribution at the single-cell level using an image sensor based on an ordinary bright-field optical microscope that divided the cell staining images into their red (R) and blue (B) channels. The grayscale of the R and B channels was subjected to a ratiometric operation to generate ratiometric grayscale cell images of the microscope. A standard curve of pH against ratiometric grayscale curve was then obtained by incubating HeLa cells at pH 6.00-7.60 in a high concentration K+ ion buffer solution containing nigericin for obtaining certain intracellular pH values. A good correlation was evidenced between pH and the ratiometric grayscale of the R and B channels in the pH range of 6.00-7.60. Subsequently, the intracellular pH value of the A549 cells under the experimental conditions was measured to be 7.22 ± 0.01 by the method. Furthermore, the changes in the intracellular pH of HeLa cells stimulated with hydrogen peroxide were sensitively monitored, which demonstrated the applicability of the method. Due to its ease of use, the developed colorimetric microscopy pH detection and monitoring method provide prospects for pH-related single-cell studies.

Development of a magneto-optical Kerr microscope using a 3D printer

Background Magneto-optical Kerr effect (MOKE) microscopes are powerful experimental tools to observe magnetic domains in magnetic materials. These devices are, however, typically large, unportable, and expensive (∼ several million yen), and therefore prevent many researchers in the field of materials science from easy access to study real-space images of magnetic domains. Methods To overcome these issues, we utilized data from “The OpenFlexure Project” developed by the University of Bath and the University of Cambridge. The purpose of this project is to make high-precision mechanical positioning of the studied sample available to anyone with a 3D printer, especially for use in microscopes. We built a low-cost and portable MOKE microscope device with a 3D printer. We redesigned the 3D modeling data of an ordinary optical microscope provided by The OpenFlexure project and incorporated additional elements, such as optical polarizers and an electromagnetic coil into the primarily designed microscope that did not originally have these elements. Results We successfully observed magnetic domains and their real-space motions induced by magnetic fields using the palm-sized low-cost MOKE microscope, which costs approximately 30,000 yen in raw materials to construct. Conclusions Our methodology to assemble a low-cost MOKE microscope will enable researchers working in the field of materials science to observe magnetic domains more easily without commercial equipment.

Pelatihan Pengembangan Mikroskop Digital sebagai Media Pembelajaran di SMAN 3 Kefamenanu

A microscope is a very important tool in learning biology. The preparation media cannot be used optimally if it is not supported by a microscope. Technological developments have succeeded in changing ordinary light microscopes into digital ones. Adding a camera to a microscope and installing software on a computer becomes an effective and efficient biology learning medium. This activity aims to provide training to teachers and students regarding the development of digital microscopes as learning media at SMAN 3 Kefamenanu. The methods used in this service activity include lectures, direct observation practice and group discussions. The results obtained from this activity are that teachers and students gain knowledge about digital microscopes and preparations, are able to operate digital microscopes and make fresh preparations independently. The evaluation results showed that 85% of participants were very satisfied with the service activities that had been carried out. Training on developing digital microscopes as a learning medium at SMAN 3 Kefamenanu can have a positive influence. Teachers and students are greatly helped in practicum-based biology learning activities. Practical activities can take place effectively and efficiently. Apart from that, teachers and students also gain knowledge and experience regarding digital microscopes, operating digital microscopes, calibration, and making fresh preparations.

Insight into the effect of microwave treatment on fat loss, fatty acid composition and microstructure of pork subcutaneous back fat

In this study, the effects of different microwave treatment times (10, 20, 30, 40, 50, and 60 s) on fat loss, fatty acid profile, lipid transfers, and microstructure of pork subcutaneous back fat (PSBF) were investigated. Results showed that the fat loss of PSBF after microwave treatment for 60 s was significantly increased (53.31%). Microwave treatment increased the proportion of saturated fatty acids but reduced the relative content of monounsaturated and polyunsaturated fatty acids. Cryo-scanning electron microscopy, ordinary optical microscope and confocal laser scanning microscope analysis demonstrated that microwave treatment facilitated a large outflow of fat from the adipocytes, and the increasing and then decreasing changes in the size of individual adipocytes. Magnetic resonance imaging enabled the visualization of fat migration of PSBF. Overall, PSBF can be used as a biological model to evaluate the effect of microwave treatment on the change and migration mechanism of fat.

Strain-induced damage analysis of hyperelastic material through scanning electron microscopy: A statistical approach using digital image processing

In order to acquire topographic/morphological images with superior resolution and depth of focus than an ordinary optical microscope, scanning electron microscopy (SEM) is used to take photographs of a surface of materials or specimens at a desired point. In the context of polymer and rubber materials science, SEM investigations frequently try to visualize phase morphology, surface and cross-sectional topography, and surface molecular order, and clarify damage mechanisms. In the case of rubber vulcanizates, test specimens are frequently put through a variety of mechanical property assessment procedures, including tensile, flexing, fatigue, abrasion, and tear tests, in order to finalize compounding parameters for required levels of quality. The damaged surface of the test specimens exhibits distinctive topographical characteristics, which are captured as an SEM picture and associated with relevant strength properties. However, the majority of the time, the relationship between strength attributes and the type of surface topography is qualitative. Surface roughness measurement metrics such as root mean square (r.m.s) roughness, average roughness, and peak-to-valley distance are frequently determined for quantitative research using standard software accessible in an SEM. The primary goal of this research is to use a statistical/spectral-based technique for quantitatively measuring surface topography using SEM. The Mullins stress-softening phenomenon of an isotropic, incompressible, hyperelastic rubberlike material is predicted using a phenomenological model. A simple exponential damage function characterizes the model, which represents deformation-induced microstructural degradation of rubberlike material.

Development of a magneto-optical Kerr microscope using a 3D printer

Background: Magneto-optical Kerr effect (MOKE) microscopes are powerful experimental tools to observe magnetic domains in magnetic materials. These devices are, however, typically large, unportable, and expensive (∼ several million yen), and therefore prevent many researchers in the field of materials science from easy access to study real-space images of magnetic domains. Methods: To overcome these issues, we utilize data from ”The OpenFlexure Project” developed by the University of Bath and the University of Cambridge. The purpose of this project is to make high-precision mechanical positioning of the studied sample available to anyone with a 3D printer, especially for use in microscopes. We built a low-cost and portable MOKE microscope device by a 3D printer. We redesigned the 3D modeling data of an ordinary optical microscope provided by The OpenFlexure project and incorporated additional elements such as optical polarizers and an electro-magnetic coil into the primarily designed microscope that did not originally have these elements. Results: We successfully observed magnetic domains and their real-space motions induced by magnetic fields using the palm-sized low-cost MOKE microscope, which costs approximately 20,000 yen in raw materials to construct. Conclusions: Our methodology to assemble a low-cost MOKE microscope will enable researchers working in the field of materials science to more easily observe magnetic domains without commercial equipment.

A multi-aspect analysis of two analogous aspergillus spp. belonging to section Flavi: aspergillus flavus and aspergillus oryzae

Microfungal isolates were routinely identified depending on both macro and micro morphological characteristics, sometimes, some fungal isolates appeared to be similar and such cases caused severe confusion for mycologists during the preliminary identification. During our previous studies dealing with isolation of fungi for some biotechnological applications; two mystifying species Aspergillus flavus and Aspergillus oryzae showed similar cultural and macroscopic features. Therefore, the current study aimed to easily distinguish between these two species depending on simple approaches which are routinely followed by a large segment of researchers. Investigation of the macroscopic features was performed to check the fungal growth on four different media (PDA, MEA, YES, and CYA) followed by microscopic examination using an ordinary light microscope, and scanning electron microscope SEM. Also, screening of secondary metabolites for both strains was preliminarily identified to find out the difference between their metabolic profiles. Finally, ITS rDNA was involved to clarify the molecular differences along their partial sequence. Conclusively, the BLAST strategy confirmed the similarity of ITS rDNA segments of both fungal strains that supported our hypothesis. The color of the fungal growth is a very critical factor whereas it is extensively influenced by the type of cultivation media. Accordingly, the YES medium was an inspiring tool assisting in prompt differentiation during the culture investigation step whereas A. oryzae and A. flavus appeared significant mustard yellow and olive green respectively. During the microscopic examination, the CYA medium also had a robust effect on the formation of the conidial chain whereas the knit long chain was observed in A. oryzae while the conidia appeared scattered and not in a chain in the case of A. flavus. Likewise, both two strains possessed different metabolic profiles where A. oryzae is not an Afla toxin producer, unlike A. flavus.

New technology offers quick and superior optical characterizations of micron-scale systems

A simple, compact, and low-cost instrument turns an ordinary microscope into a fast, accurate thin-film analyzer.

Damage-free LED lithography for atomically thin 2D material devices

Desired electrode patterning on two-dimensional (2D) materials is a foremost step for realizing the full potentials of 2D materials in electronic devices. Here, we introduce an approach for damage-free, on-demand manufacturing of 2D material devices using light-emitting diode (LED) lithography. The advantage of this method lies in mild photolithography by simply combining an ordinary optical microscope with a commercially available LED projector; the low-energy red component is utilized for optical characterization and alignment of devices, whereas the high-energy blue component is utilized for photoresist exposure and development of personal computer designed electrode patterns. This method offers maskless, damage-free photolithography, which is particularly suitable for 2D materials that are sensitive to conventional lithography. We applied this LED lithography to device fabrication of selected nanosheets (MoS2, graphene oxides and RuO2), and achieved damage-free lithography of various patterned electrodes with feature sizes as small as 1–2 μm. The LED lithography offers a useful approach for cost-effective mild lithography without any costly instruments, high vacuum, or complex operation.

Observation of the protein expression level via naked eye: Pt clusters catalyze non-color molecules into brown-colored molecules in cells.

α v β 3 is overexpressed in various tumor cells and plays a key role in tumor genesis, invasion, and metastasis. Therefore, it is of great significance to precisely detect the α v β 3 level in cells via a simple method. For this purpose, we have constructed a peptide-coated platinum (Pt) cluster. Due to its bright fluorescence, well-defined Pt atom numbers, and peroxidase-like catalytic activity, this cluster can be used to evaluate α v β 3 levels in cells by fluorescence imaging, inductively coupled plasma mass spectrometry (ICP-MS), and catalytic amplification of visual dyes, respectively. In this report, the expression level of α v β 3 in living cells is well-detected by the naked eye under an ordinary light microscope when the Pt cluster binds to αvβ3 in cells and catalyzes non-color 3,3'-diaminobenzidine (DAB) into brown-colored molecules in situ. Moreover, SiHa, HeLa, and 16HBE cell lines with different α v β 3 expression levels can be visually distinguished by the peroxidase-like Pt clusters. This research will provide a reliable method for the simple detection of α v β 3 levels in cells.

Single Molecule Imaging with Liquid Phase Electron Microscopy

AbstractFew single‐molecule experiments have enabled the direct imaging of functional biomacromolecules in real‐time in their native liquid environments, resolving their conformational adaptations, transient interactions, and intermediate states. Liquid phase electron microscopy (LP‐EM), due to its unique combination of spatial and temporal resolution, has shown to be a promising tool. Recent experiments have enabled successful imaging of intact structures of organic molecules and biological systems with an ordinary electron microscope. Adapting image processing methods and quantitative data analysis from single particle experiments based on the optical microscope, quantifying motion and relaxation of these interacting molecules allows the experimental observations of pathways, to test theoretical predictions, and discovery of new mechanisms. Combining LP‐EM with tomography, fluorescence, and mass spectroscopy allows for probing multi‐dimensional structural and dynamic information. Challenges remain in obtaining high‐quality data in large quantities, which can be improved by developing new liquid cell platforms and machine learning‐based data analysis.

Single-shot spatial light interference microscopy by demultiplexing based on polarization gratings

Off-axis interferometric modules built on an ordinary bright field microscope make it possible to achieve single-shot quantitative phase imaging (QPI) by adding sufficient spatial carrier into the interferograms. However, compared with its on-axis counterparts, imaging configurations for off-axis interferometric modules have several disadvantages regarding optical aberration, stability, and space-bandwidth utilization of the lenses system. Herein, by demultiplexing technology based on polarization gratings, we propose a single-shot spatial light interference microscopy named as polarization-multiplexing light interference microscopy (PLIM) with on-axis imaging configuration to realize single-shot QPI. Although the imaging system is on-axis, the PLIM system still can generate sufficient spatial carrier, so we can adjust the orientation and absolute value of the spatial carrier independently of the magnification ratio of the imaging system. The experimental results prove that the PLIM system has better temporal phase stability compared with conventional grating-based QPI technologies and is suitable for high resolution QPI.

Role of anti-Giardia recombinant cyst wall protein IgG polyclonal antibodies in diagnosis and protection

Giardia duodenalis (G. duodenalis) is an infectious protozoan that has a global distribution especially in the hot climate. Around 200 million people are infected worldwide annually by Giardia, but infection is not always accompanied by symptoms, especially in endemic countries. Using traditional microscopy techniques in diagnosis, both in stool and water samples were less sensitive when compared to immunological methods; and the need for new diagnostic methods was necessary. Also, protection from infection is required in endemic areas. Therefore, the study aimed to produce anti- G. duodenalis IgG polyclonal antibodies (pAbs) by immunizing rabbit by G. duodenalis cyst recombinant protein. The produced antibodies were evaluated in the detection of G. duodenalis antigens in patients’ stool and water samples from endemic areas across River Nile; where pAbs were used as a coating and a peroxidase conjugate antibody in sandwich ELISA. Moreover, pAbs were tested for the protection of mice from giardiasis. Sandwich ELISA using pAb has succeeded in the detection of G. duodenalis coproantigens in stool samples by a sensitivity of 97% and a specificity of 92.72%. Moreover, G. duodenalis cyst was detected in only seven water samples by ordinary microscopy; while sandwich ELISA revealed nineteen positive results. IgG pAb (1/200 µg/ml) protected mice from giardiasis; which was evident from the reduction in cysts and trophozoites numbers. We recommended the use of sandwich ELISA to monitor water quality, investigate environmental contamination and diagnosis in patients' stools. The pAbs can be prepared in large amount and used in field diagnosis and protection. This will help in the early diagnosis of G. duodenalis in water, which in turn can control outbreaks in rural areas.

Scanning Electron Microscopic Observation of Endosperm Chalkiness Traits of OsAAP1 Mutant in Rice Gains

In order to investigate the effect of OsAAP1 gene mutation on chalkiness traits of rice endosperm and the microstructure of starch granules in endosperm, we used rice OsAAP1 mutants as experimental materials, with the help of ordinary optical microscopes and scanning electron microscopes, the morphological structure and arrangement of chalkiness traits and starch granules in the endosperm of Zhonghua 11 rice were observed in rice OsAAP1 mutants and controls. The results showed that the chalkiness rate, chalkiness area and chalkiness degree in rice endosperm of the OsAAP1 mutants were significantly increased, and the morphological structure, arrangement and development of chalkiness traits and starch granules were closely related. Compared with the control Zhonghua 11, there were also significant differences in the shape, structure and arrangement of starch granules between chalky rice and non-chalky rice. However, there was no significant difference between the starch granules in the transparent part of chalky rice and non-chalky rice, and the starch granules in the areas with low chalkiness degree developed well. Thus, the OsAAP1 mutation of rice could significantly increase the chalkiness traits of rice endosperm, and have an important impact on the morphology, structure and development of starch granules in its endosperm.

Experimental Guidelines to Image Transient Single-Molecule Events Using Graphene Liquid Cell Electron Microscopy.

In quest of the holy grail to "see" how individual molecules interact in liquid environments, single-molecule imaging methods now include liquid-phase electron microscopy, whose resolution can be nanometers in space and several frames per second in time using an ordinary electron microscope that is routinely available to many researchers. However, with the current state of the art, protocols that sound similar to those described in the literature lead to outcomes that can differ. The key challenge is to achieve sample contrast under a safe electron dose within a frame rate adequate to capture the molecular process. Here, we present such examples from different systems─synthetic polymer, lipid assembly, DNA-enzyme─in which we have done this using graphene liquid cells. We describe detailed experimental procedures and share empirical experience for conducting successful experiments, starting from fabrication of a graphene liquid cell, to identification of high-quality liquid pockets from desirable shapes and sizes, to effective searching for target sample pockets under electron microscopy, and to discrimination of sample molecules and molecular processes of interest. These experimental tips can assist others who wish to make use of this method.

High-resolution polarization-sensitive Fourier ptychography microscopy using a high numerical aperture dome illuminator.

Polarization-sensitive Fourier-ptychography microscopy (pFPM) allows for high resolution imaging while maintaining a large field of view, and without mechanical movements of optical-setup components. In contrast to ordinary light microscopes, pFPM provides quantitative absorption and phase information, for complex and birefringent specimens, with high resolution across a wide field of view. Using a semi-spherical home-built LED illumination array, a single polarizer, and a 10x /0.28NA objective, we experimentally demonstrate high performance pFPM with a synthesized NA of 1.1. Applying the standard quantitative method, a measured half-pitch resolution of 244 nm is achieved for the 1951 USAF resolution test target. As application examples, the polarimetric properties of a herbaceous flowering plant and the metastatic carcinoma of human liver cells are analyzed and quantitatively imaged.

Revealing nanostructures in brain tissue via protein decrowding by iterative expansion microscopy.

Many crowded biomolecular structures in cells and tissues are inaccessible to labelling antibodies. To understand how proteins within these structures are arranged with nanoscale precision therefore requires that these structures be decrowded before labelling. Here we show that an iterative variant of expansion microscopy (the permeation of cells and tissues by a swellable hydrogel followed by isotropic hydrogel expansion, to allow for enhanced imaging resolution with ordinary microscopes) enables the imaging of nanostructures in expanded yet otherwise intact tissues at a resolution of about 20 nm. The method, which we named 'expansion revealing' and validated with DNA-probe-based super-resolution microscopy, involves gel-anchoring reagents and the embedding, expansion and re-embedding of the sample in homogeneous swellable hydrogels. Expansion revealing enabled us to use confocal microscopy to image the alignment of pre-synaptic calcium channels with post-synaptic scaffolding proteins in intact brain circuits, and to uncover periodic amyloid nanoclusters containing ion-channel proteins in brain tissue from a mouse model of Alzheimer's disease. Expansion revealing will enable the further discovery of previously unseen nanostructures within cells and tissues.

Nondestructive visualization of graphene on Pt with methylene blue surface modification

Efficient large-scale nondestructive quality assessment of graphene on Pt is essential to the in-depth growth research and practical applications of graphene. Here, we present a very simple method for directly observing the domains and defects in graphene on Pt using an ordinary optical microscope. This was achieved by modifying graphene on Pt using methylene blue (MB). Because the chemical activities of graphene and Pt surface differ significantly, the adsorption and reaction of MB on graphene and platinum surface differ. We can determine the distribution of graphene crystal domains and defects by comparing the colors in the optical images. In addition, this characterization method causes no obvious damage to the Pt substrate and graphene. Moreover, it does not affect the recycling of the substrate or the subsequent characterization or application of graphene. Our study provides a nondestructive method for measuring the quality of graphene on Pt on a large scale, as well as a reference for the characterization and doping of other two-dimensional materials.

Elaboration, characterization and applications of SnO2, 2 %Gd-SnO2 and 2 %Gd-9 %F-SnO2 thin films for the photocatalytic degradation of MB by USP method

• An investigation on SnO 2 , 2 %Gd-SnO 2 and 2 %Gd-9 %F-SnO 2 thin films have been synthesized by USP method. • Effect of doping and co-doping on the structrales, microdtructurales, optical and electrical properties of SnO 2 thin films. • The development of the parameters: lattice parameters, average crystalline sizes, Seeback coefficient and energy gap with doping and co-doping are discussif. • The photocatalytic degradation of methylene blue was studied using 2 %Gd-SnO 2 and 2 %Gd-9 %F-SnO 2 thin films. In this study, SnO 2 , 2 %Gd-SnO 2 , and 2 %Gd-9 %F-SnO 2 films were grown from tin chloride solutions by the ultrasonic spray technique on ordinary microscope glass substrates heated to a fixed temperature of 450 °C for photocatalytic application. The microstructural characterization of the thin films obtained was performed using X-ray diffraction (XRD), SEM, EDAX, and Infrared Spectroscopy (IR). The optical properties of thin films were studied by UV–Visible spectroscopy. To examine the electrical properties, we use the four-point method, the Seebeck effect and the Mott Shottky analysis. X-ray difraction reveals the formation of the routile phase of SnO 2 in all thin films with average crystallite size estimated in the range of 24–62 nm. The co-doping in (Gd-F) permitted an improvement of the thermoelectric properties of SnO 2 . Moreover, we have demonstrated that an increase in grain size allows toimprove the Seebeck coefficient, to increase the electrical conductivity, on the other hand the doping in (Gd), presents a decrease in the size of the crystallites, which improves the degradation of methyl blue. The UV–Visible spectra of optical transmission of the layers show that the pure, doped and co-doped SnO2 films are of good quality with a transmittance of 73–83% in the visible and have an optical gap evaluated between 3.89, 3.98 and 3.99 eV respectively.