This is the third article within a three-part series on Fourier ptychography, which is a computational microscopy technique for high-resolution, large field-of-view imaging. While the previous articles introduced the working principles of the technique, in this article we focus on the practical benefits that it brings to the imaging community. We present a didactic overview of the most important and well-established practical use-cases such as gigapixel imaging, quantitative phase contrast, thick sample imaging, and aberration metrology. We also discuss how Fourier ptychography can leave the visible light domain and venture into the realm of smaller wavelengths such as X-rays and electrons, among other topics.

In this study, the gut structure and excretory system of Carpocoris mediterraneus which is phytophagous insect, were described with light and electron microscopies and discussed in relation to other Heteroptera species. The salivary system has two principal and accessory salivary glands, two principal and accessory gland ducts. The salivary gland and duct wall have a single layer of cuboidal cells. The duct lumen is surrounded by a thick intima layer. In the cytoplasm of the epithelial cells are seen vesicles. The gut includes fore, mid, and hindguts. The foregut consists of a long narrow tubular pharynx which opens into a slightly wider esophagus. The esophagus is thin walled and in turn opens into the midgut. The midgut has four regions (V1–V4). V1–V4 walls have a monolayered epithelium. V1 epithelium is double-nucleated. V1 cytoplasm contains numerous vesicles, secretory granules, spherocrystals, and cytoplasmic inclusions. Rod-shaped bacteria are seen in V4 lumen. The hindgut has pylorus and rectum. Malpighian tubules were attached in the pylorus. Malpighian tubules have a single-layer cuboidal epithelium. In their lumen, there are spherocrystals. The rectum wall has a monolayer of squamous epithelium and muscle layer. Numerous bacteria and uric acid crystals are seen in its lumen.

This research reports for the first time the anatomical characteristics of all species belonging to Baccharis subgenus Coridifoliae (Asteraceae). The anatomy and micro-morphology of aerial vegetative organs of ten species: B. albilanosa, B. artemisioides, B. bicolor, B. coridifolia, B. erigeroides, B. napaea, B. ochracea, B. pluricapitulata, B. scabrifolia, and B. suberectifolia are investigated by light and scanning electron microscopy. The number of secretory ducts, crystal morphology, presence or absence of conical nonglandular trichomes, leaves cross-section shape, margin morphology, anticlinal epidermal cell walls shape, and cuticle structure were identified as characters with diagnostic value for species. Similarity cluster analysis allows the formation of three groups based on a percentage of similarity between 45 and 84%. Some species showed differential characteristics as the presence of up to four secretory ducts in the midrib in B. albilanosa; smooth cuticles onboth sides of the leaf epidermis in B. erigeroides; flat midrib shape on both sides of the leaves in B. napaea; and convex–flat midrib shape in B. suberectifolia. The remaining species can be differentiated by a set of anatomical features. Anatomical and histochemical characteristics of stems and leaves provided data to support species identification.

The in vitro models are receiving growing attention in studies on skin permeation, penetration, and irritancy, especially for the preclinical development of new transcutaneous drugs. However, synthetic membranes or cell cultures are unable to effectively mimic the permeability and absorption features of the cutaneous barrier. The use of explanted skin samples maintained in a fluid dynamic environment would make it possible for an in vitro experimentation closer to in vivo physiological conditions. To this aim, in the present study, we have modified a bioreactor designed for cell culture to host explanted skin samples. The preservation of the skin was evaluated by combining light, transmission, and scanning electron microscopy, for the histo/cytological characterization, with nuclear magnetic resonance spectroscopy, for the identification in the culture medium of metabolites indicative of the functional state of the explants. Our morphological and metabolomics results demonstrated that fluid dynamic conditions ameliorate significantly the structural and functional preservation of skin explants in comparison with conventional culture conditions. Our in vitro system is, therefore, reliable to test novel therapeutic agents intended for transdermal administration in skin samples from biopsies or surgical materials, providing predictive information suitable for focused in vivo research and reducing animal experimentation.

Metchnikovellids are a deep-branching group of microsporidia, parasites of gregarines inhabiting the alimentary tract of polychaetes and some other invertebrates. The diversity and phylogeny of these hyperparasites remain poorly studied. Modern descriptions and molecular data are still lacking for many species. The results of a light microscopy study and molecular data for Metchnikovella spiralis Sokolova et al., 2014, a hyperparasite of the eugregarine Polyrhabdina sp., isolated from the polychaete Pygospio elegans, were obtained. The original description of M. spiralis was based primarily on the analysis of stained preparations and transmission electron microscopy images. Here, the species description was complemented with the results of in vivo observations and phylogenetic analysis based on the SSU rRNA gene. It was shown that in this species, free sporogony precedes sac-bound sporogony, as it occurs in the life cycle of most other metchnikovellids. Spore sacs are entwined with spirally wound cords, and possess only one polar plug. Phylogenetic analyses did not group M. spiralis with M. incurvata, another metchnikovellid from the same gregarine species, but placed it as a sister branch to Amphiacantha. The paraphyletic nature of the genus Metchnikovella was discussed. The taxonomic summary for M. spiralis was emended.

Avian malaria is a mosquito-borne disease caused by Plasmodium spp. protozoa. Although these parasites have been extensively studied in North America and Eurasia, knowledge on the diversity of Plasmodium, its vectors and avian hosts in Africa is scarce. In this study, we report on natural malarial infections in free-ranging sparrows (Passer domesticus) sampled at Giza Governorate, Egypt. Parasites were morphologically characterized as Plasmodium cathemerium based on the examination of thin blood smears from the avian host. Sequencing a fragment of the mitochondrial cytochrome b gene showed that the parasite corresponded to lineage PADOM02. Phylogenetic analysis showed that this parasite is closely related to the lineages SERAU01 and PADOM09, both of which are attributed to P. cathemerium. Experimental infection of Culex pipiens complex was successful, with ookinetes first detected at 1-day post infection (dpi), oocysts at 4 dpi and sporozoites at 6 dpi. The massive infection of the salivary glands by sporozoites corroborates that Cx. pipiens complex is a competent vector of PADOM02. Our findings confirm that Plasmodium lineage PADOM02 infects sparrows in urban areas along the Nile River, Egypt, and corroborate that Cx. pipiens complex is a highly competent vector for these parasites. Furthermore, our results demonstrate that this lineage corresponds to the morphospecies P. cathemerium and not P. relictum as previously believed.

A method was developed to expedite the imaging of diatoms by optimizing their orientation. This quick technique puts grooves into a SEM stub so that as the diatoms are laid down they either land on the flat surface or in a variety of positions in the grooves, thereby reducing time required to capture taxonomically critical features.

The bioaccessibility of fat has implications for satiety and postprandial lipidaemia. The prevailing view holds that the integrity of plant cell wall structure is the primary determinant of energy and nutrient extraction from plant cells as they pass through the gastrointestinal (GI) tract. However, comparisons across nuts (walnuts, almonds and pistachios) with varying physical properties do not support this view. In the present study, masticated samples of three nuts from healthy adults were exposed to a static model of gastric digestion followed by simulated intestinal digestion. Primary outcomes were particle size and lipid release at each phase of digestion. Walnuts produced a significantly larger particle size post-mastication compared with almonds. Under gastric and intestinal conditions, the particle size was larger for walnuts compared with pistachios and almonds (P < 0·05). However, the masticated and digesta particle sizes were not related to the integrity of cell walls or lipid release. The total lipid release was comparable between nuts after the in vitro intestinal phase (P > 0·05). Microstructural examination showed ruptured and fissured cell walls that would allow digestion of cellular contents, and this may be governed by internal cellular properties such as oil body state. Furthermore, the cell walls of walnuts tend to rupture rather than separate and as walnut tissue passes through the GI tract, lipids tend to coalesce reducing digestion efficiency.

Individual plant stomata were studied within a sealed environmental chamber that encloses a volume of space around a microscope objective. A leaf that is attached to a plant is secured to a temperature-controlled microscope stage, and the top surface of the leaf is illuminated via the microscope's light source while stomata on the underside of the leaf are observed through a microscope objective. Stomatal responses are observed on a leaf exposed to environmental conditions including various illuminations and changes in ambient CO2 level, at a given temperature.

In September 2018, Australia hosted the 19th International Microscopy Congress in Sydney. As part of this conference, more than 600 teachers and students from 19 classes across the state of NSW were hosted on-site at the IMC19 Outreach Learning Space. The program resulted from a year-long collaborative effort between Australian Microscopy and Microanalysis and Microscopy Australia volunteers, academics, microscopy industry vendors, conference coordinators, STEM secondary school teachers, and local attractions. A unique feature of the program was the incorporation of the Australian national curriculum into the program via teacher-designed prework and postwork, helping to contribute to an award-winning conference.

Routine system checks are essential for supervising the performance of an advanced light microscope. Recording and evaluating the point spread function (PSF) of a given system provides information about the resolution and imaging. We compared the performance of fluorescent and gold beads for PSF recordings. We then combined the open-source evaluation software PSFj with a newly developed KNIME pipeline named PSFtracker to create a standardized workflow to track a system's performance over several measurements and thus over long time periods. PSFtracker produces example images of recorded PSFs, plots full-width-half-maximum (FWHM) measurements over time and creates an html file which embeds the images and plots, together with a table of results. Changes of the PSF over time are thus easily spotted, either in FWHM plots or in the time series of bead images which allows recognition of aberrations in the shape of the PSF. The html file, viewed in a local browser or uploaded on the web, therefore provides intuitive visualization of the state of the PSF over time. In addition, uploading of the html file on the web allows other microscopists to compare such data with their own.

Stomata (singular, “stoma”) are tiny pores through which plants breathe. Stomata are found on the upper and lower sides of leaves, on flower petals, on stems, and on roots. Scientists survey plant surfaces to determine the density and size of stomata and relate these findings to properties of the environment, such as temperature and the amounts of sunlight, humidity, oxygen, and carbon dioxide in the air when a leaf is formed. Stomata of various plants are suitable subjects for classroom laboratory activities since they may be examined by light microscopy.

Atomic force microscopy (AFM) is used in a wide range of applications for imaging sub-nanometer resolution topography, as well as for analysis of physical characteristics (such as surface stiffness and electromagnetic properties). While surface imaging and characterization on the nanoscale is of great value and is widely used across all fields of science, correlation with other types of microscopy can greatly enhance the value of those measurements. Here, we discuss a new approach that uses a small, ultra-thin AFM, which allows it to be integrated with standard light microscopes. The authors believe the design will allow for a new paradigm in microscopy, by enabling all the advantages of AFM to be combined with the advanced imaging capabilities of research-grade light microscopes.

The U.S. Patent and Trademark Office (PTO) encourages independent inventors who are not patent professionals to file and prosecute their own patent applications. Self-help books are useful guides for navigating through the PTO’s many laws and rules. When an inventor has an idea, he should document it, have the document witnessed, search all public records for novelty and unobviousness, and finally file an application with the PTO. The inventor defends his idea with an Examiner from the PTO and, if all requirements are met, receives a patent that grants the inventor a monopoly to make, use, and sell his invention for a period of time.

Artemia sp. (brine shrimp) can be used for pedagogic purposes as an instructive model organism to demonstrate the development and maturation of small marine crustaceans within a time window suited to classroom experiments. This article describes some characteristics of Artemia and gives several hints for culturing, light microscopy observations, and photomicrography. The morphological variations and successive differentiation associated with the maturation process are presented through various types of light microscopy images. Increasing differentiations of movement patterns are considered. Moreover, some suggestions of further experiments are reported, which can be used as introductions to fundamental principles of scientific work.

At present, there are no medicinal plant extracts currently available for treatment and control of fasciolosis. The present work could provide, for the first study, conclusions on the in vitro fasciolicidal properties of the ethanol extract of Terminalia catappa L. (TcCE) leaves against adult Fasciola gigantica after incubation with RPMI-1640 medium containing the TcCE at various concentrations and times when compared with triclabendazole (TCZ). The relative motility and survival index values of the TcCE-treated flukes decreased at a more rapid rate than the TCZ-treated flukes. The death of the parasites was observed after exposed to TcCE at 3 h incubation with 400, 800 and 1000 µg mL−1, and at 6 h incubation in 100 and 200 µg mL−1. Vacuolization, blebbings and partial disruption on the parasites’ tegument were observed by light microscopy. When examined by scanning electron microscopy, TcCE caused similar tegumental alterations in the parasites as those observed in TCZ treatment but with larger damage at comparative incubation periods, consisting of swelling, blebbing, disrupted blebs, loss of spines, leading to the erosion, lesion and eventual disruption of the total tegument. Therefore, the TcCE may exert its fasciolicidal effect against F. gigantica by initially causing the tegumental alteration.

A test of the qualities of polarizing filters was performed for a set of specimens including a bulk Nicol prism, standard polaroids, and special polyvinyl alcohol (PVA)-iodine thin-film filters coated on both sides by vertically oriented carbon nanotubes. The residual transmission of polarizing filters depending on the incidence angle of polarized light was examined in detail. The superior quality of polarizing film filters treated with carbon nanotubes was found. This fact allows us to propose a new application for polarizing films with carbon nanotubes for a polarizing cover glass. In such a way the cover glass may serve as an analyzer in a light polarizing microscope. Some features of optical scheme arrangement for the polarizing technique are discussed. The polarizing cover glass allows elimination of depolarization of light, which is inserted in a microscope objective. Test results of the proposed polarizing technique attest to the efficiency of using the polarizing cover glass. The new scheme for polaroid arrangement shows image-contrast enhancement by several percent in comparison with the standard layout.

The aim of the present work was to investigate birefringence and morphology of the secretory-stage enamel organic extracellular matrix (EOECM), and structural and mechanical properties of mature enamel of upper incisors from adult rats that had been treated with pamidronate disodium (0.5 mg/kg/week for 56 days), using transmitted polarizing and bright-field light microscopies (TPLM and BFLM), energy-dispersive X-ray (EDX) analysis, scanning electron microscopy (SEM) and microhardness testing. BFLM showed no morphological changes of the EOECM in pamidronate and control groups, but TPLM revealed a statistically significant reduction in optical retardation values of birefringence brightness of pamidronate-treated rats when compared with control animals (p<0.01). EDX analysis showed that pamidronate-induced statistically significant decrease in phosphorus’ quantity in outer mature enamel (p<0.01) and an increase in the calcium/phosphorus ratio in that structure (p<0.05). Pamidronate did not induce ultra-structural alterations in mature enamel as revealed by SEM and did not cause a reduction in its microhardness (p>0.05). The present study indicates that pamidronate can affect birefringence of the secretory-stage EOECM, which does not seem to be associated with significant changes in morphological and/or mechanical properties of mature enamel.

Presently, no effective anthelmintic drugs have been used to treat and control paramphistomosis, a severe disease of ruminants. In this study, we have investigated the in vitro anthelmintic effect of the leaves of Terminalia catappa L. crude extract (TcCE) and albendazole (ABZ) on adult Fischoederius cobboldi after incubating the flukes in RPMI-1640 medium containing the TcCE at various doses and times. The TcCE-treated flukes at all dosages exhibited rapid decrease of motility, and the relative motility (RM) values were decreased sharply from start to 3 h. Worms were killed after 6 and 12 h of treatment with 1000, 1500 and 2000 µg mL−1 as well as 500 µg mL−1 of TcCE, respectively. By light microscopy examination, the flukes exhibited the earliest alteration in a limited area of the tegument. At scanning electron microscopy level, the flukes’ tegument showed similar sequence of morphological alterations after treatment with ABZ and TcCE that consisted of swelling of ridges and folds, followed by blebbing and rupturing of the blebs, leading to the erosion, lesion and disruption of the tegument. Hence, in vivo studies should be performed to examine whether the TcCE may serve as a powerful anthelmintic drug for treatment of paramphistomosis.

This work evaluates the use of light microscopes (LMs) as a tool for interlaminar fracture of polymer composite investigation with the aid of correlative fractography. Correlative fractography consists of an association of the extended depth of focus (EDF) method, based on reflected LM, with scanning electron microscopy (SEM) to evaluate interlaminar fractures. The use of these combined techniques is exemplified here for the mode I fracture of carbon–epoxy plain-weave reinforced composite. The EDF-LM is a digital image-processing method that consists of the extraction of in-focus pixels for each x-y coordinate in an image from a stack of Z-ordered digital pictures from an LM, resulting in a fully focused picture and a height elevation map for each stack. SEM is the most used tool for the identification of fracture mechanisms in a qualitative approach, with the combined advantages of a large focus depth and fine lateral resolution. However, LMs, with EDF software, may bypass the restriction on focus depth and present enough lateral resolution at low magnification. Finally, correlative fractography can provide the general comprehension of fracture processes, with the benefits of the association of different resolution scales and contrast modes.