Imaging technologies have played a pivotal role in advancing biological research by enabling visualization of biological structures and processes. While traditional electron microscopy (EM) produces two-dimensional images, emerging techniques now allow high-resolution three-dimensional (3D) characterization of specimens in situ, meeting growing needs in molecular and cellular biology. Combining transmission electron microscopy (TEM) with serial sectioning inaugurated 3D imaging, attracting biologists seeking to explore cell ultrastructure and driving advancement of 3D EM reconstruction. By comprehensively and precisely rendering internal structure and distribution, 3D TEM reconstruction provides unparalleled ultrastructural insights into cells and molecules, holding tremendous value for elucidating structure-function relationships and broadly propelling structural biology. Here, we first introduce the principle of 3D reconstruction of cells and tissues by classical approaches in TEM and then discuss modern technologies utilizing TEM and on new SEM-based as well as cryo-electron microscope (cryo-EM) techniques. 3D reconstruction techniques from serial sections, electron tomography (ET), and the recent single-particle analysis (SPA) are examined; the focused ion beam scanning electron microscopy (FIB-SEM), the serial block-face scanning electron microscopy (SBF-SEM), and automatic tape-collecting lathe ultramicrotome (ATUM-SEM) for 3D reconstruction of large volumes are discussed. Finally, we review the challenges and development prospects of these technologies in life science. It aims to provide an informative reference for biological researchers.

A literature review for a recent ultrastructural study of a trichinelloid eggshell revealed consistently occurring errors in the literature on nematode eggshell anatomy. Examples included nematodes of medical, veterinary, and agricultural importance in several orders. Previous researchers had warned of some of these errors decades ago, but a comprehensive solution was not offered until 2012 when a clarifying new anatomical and developmental interpretation of nematode eggshells was proposed by members of the Caenorhabditis elegans Research Community. However, their findings were explained using arcane acronyms and technical jargon intended for an audience of experimental molecular geneticists, and so their papers have rarely been cited outside the C. elegans community. Herein we (1) provide a critical review of nematode eggshell literature in which we correct errors and relabel imagery in important historical reports; (2) describe common reporting errors and their causes using language familiar to researchers having a basic understanding of microscopy and nematode eggs; (3) recommend a new hexalaminar anatomical and terminological framework for nematode eggshells based on the 2012 C. elegans framework; and (4) recommend new unambiguous terms appropriate for the embryonated/larvated eggs regularly encountered by practicing nematodologists to replace ambiguous or ontogenetically restricted terms in the 2012 C. elegans framework. We also (5) propose a resolution to conflicting claims made by the C. elegans team versus classical literature regarding Layer #3, (6) extend the C. elegans hexalaminar framework to include the polar plugs of trichinelloids, and (7) report new findings regarding trichinelloid eggshell structure.
UNASSIGNED: La coque des œufs des nématodes : un nouveau cadre anatomique et terminologique, avec une revue critique de la littérature pertinente et des lignes directrices suggérées pour l’interprétation et la communication de l’imagerie des coques des œufs.
UNASSIGNED: Une revue de la littérature pour une étude ultrastructurale récente de la coque de l’œuf d’un trichinelloïde a révélé des erreurs récurrentes dans la littérature sur l’anatomie de la coque de l’œuf des nématodes. Les exemples comprenaient des nématodes d’importance médicale, vétérinaire et agricole dans plusieurs ordres. Des chercheurs avaient mis en garde contre certaines de ces erreurs il y a des décennies, mais une solution complète n’a été proposée qu’en 2012, lorsqu’une nouvelle interprétation anatomique et développementale clarifiant la structure des coques des œufs de nématodes a été proposée par des membres de la communauté de recherche de Caenorhabditis elegans. Cependant, leurs découvertes ont été expliquées à l’aide d’acronymes mystérieux et d’un jargon technique destiné à un public de généticiens moléculaires expérimentaux, et leurs articles ont donc rarement été cités en dehors de la communauté de C. elegans. Ici, nous (1) fournissons une revue critique de la littérature sur les coques des œufs de nématodes dans laquelle nous corrigeons les erreurs et réétiquetons les images dans des rapports historiques importants; (2) décrivons les erreurs de description courantes et leurs causes en utilisant un langage familier aux chercheurs ayant une compréhension de base de la microscopie et des œufs de nématodes; (3) recommandons un nouveau cadre anatomique et terminologique hexalaminaire pour les coques des œufs de nématodes basé sur le cadre de C. elegans de 2012; et (4) recommandons de nouveaux termes non ambigus appropriés pour les œufs embryonnés/larvés régulièrement rencontrés par les spécialistes de nématodes en exercice pour remplacer les termes ambigus ou à restriction ontogénétique dans le cadre de C. elegans de 2012. Nous proposons également (5) une résolution des affirmations contradictoires de l’équipe C. elegans par rapport à la littérature classique concernant la couche 3, (6) étendons le cadre hexalaminaire de C. elegans pour inclure les bouchons polaires des trichinelloïdes, et (7) signalons de nouvelles découvertes concernant la structure de la coque des œufs des trichinelloïdes.

A 16-year-old female spayed domestic shorthaired cat was examined for lameness and a mass on the fourth digit of the right hindlimb. Cytologic examination of an aspirate of the mass revealed large discrete cells admixed with low numbers of well-granulated mast cells. The discrete cells contained single to many variably sized light pink to purple granules in their cytoplasm and had pleomorphic nuclei, with intranuclear cytoplasmic inclusions. Karyomegalic, binucleated and multinucleated cells were seen. Histologic examination of formalin-fixed sections of the excised mass showed a mildly infiltrative, unencapsulated, multinodular dermal mass that extended into the subcutis and consisted of similar discrete cells. On immunohistochemical staining, the tumor cells expressed ionized calcium-binding adapter molecule 1 (Iba1) and CD18. The tumor cells did not express CD3, CD20, CD117, pancytokeratin (AE1/AE3), melanoma antigen (Melan-A), multiple myeloma oncogene-1 (MUM1), melanoma-associated antigen (PNL-2), and S-100. Low numbers of tumor cells expressed CD204 and protein gene product 9.5 (PGP9.5). Granules were variably positive for Periodic-acid Schiff (PAS) and Alcian blue. On transmission electron microscopy, the cells contained filopodia, abundant endoplasmic reticulum, and moderate numbers of low-density membrane-bound granules. This case documents a previously undescribed granular variant of a histiocytic tumor in a cat.

Carbon quantum dots (CQDs) have emerged as one of the most promising nanomaterials in the carbon nanostructures family in recent years due to their low toxicity, simple synthetic methods, unique fluorescence emission, good photostability, excellent water solubility, high specific surface areas and outstanding electronic properties. They have thus been employed in a wide range of applications, including fluorescent sensing, electrochemical sensing, bioimaging, drug delivery, antimicrobial studies, antioxidants, and photocatalysis. CQDs drawn great interest in sensing applications due to their unique photochemical, electrochemical and electrochemiluminescence properties. They exhibit excitation wavelength-dependent or -independent photoluminescence (PL) behaviour, high quantum yield, and promising binding ability with analytes, which make them an ideal candidate for use in PL based sensing platforms. Excessive use of agrochemicals in farm fields can pollute the environment and have potentially adverse health effects on aquatic and human life. Since there are very few monitoring techniques are available for sensing such harmful substances, there is an urgent need to develop a sensor for the facile, rapid and on-site detection and quantification of agrochemical residues in the environment. Several CQD-based fluorophores for detecting agrochemical residues employing static or dynamic quenching processes have recently been published. The key quenching mechanisms involved in the sensing process include FRET, PET and IFE. The first part of this review intends to provide a comprehensive overview of various techniques to characterize CQDs such as UV-vis., FT-IR, PL, XRD, NMR, TEM, TGA, XPS and Raman analysis. In addition application of CQDs as fluorescent sensors for agrochemical residue in different media are summarized in this reiew. The LOD values and rapid action of the sensor demonstrates significant advantages of these methods over conventional analytical procedures.

Nanomaterials have been widely used in many fields in the last decades, including electronics, biomedicine, cosmetics, food processing, buildings, and aeronautics. The application of these nanomaterials in the medical field could improve diagnosis, treatment, and prevention techniques. Graphene oxide (GO), an oxidized derivative of graphene, is currently used in biotechnology and medicine for cancer treatment, drug delivery, and cellular imaging. Also, GO is characterized by various physicochemical properties, including nanoscale size, high surface area, and electrical charge. However, the toxic effect of GO on living cells and organs is a limiting factor that limits its use in the medical field. Recently, numerous studies have evaluated the biocompatibility and toxicity of GO in vivo and in vitro. In general, the severity of this nanomaterial\'s toxic effects varies according to the administration route, the dose to be administered, the method of GO synthesis, and its physicochemical properties. This review brings together studies on the method of synthesis and structure of GO, characterization techniques, and physicochemical properties. Also, we rely on the toxicity of GO in cellular models and biological systems. Moreover, we mention the general mechanism of its toxicity.

Syngnathidae (seahorses, pipefish and seadragons) are charismatic species commonly kept in commercial aquaria, but published literature on syngnathid diseases is limited and immunohistochemical techniques not routinely employed. A retrospective review of 2,541 syngnathid submissions received between March 2003 and October 2016 identified 18 neoplasms including germ cell tumours, exocrine pancreatic and intestinal carcinomas, chromatophoromas, and single cases of lymphoma, thyroid and renal carcinoma, swim bladder and pituitary adenoma. Big-bellied seahorses accounted for 19% of submissions, but 50% of neoplasms were diagnosed in this species. This study includes the first reported cases of germ cell tumours, chromatophoroma, thyroid carcinoma and pituitary adenoma in Syngnathidae and the first reports of neoplasia in pipefish species. Out of nine commercial antibodies trialled for immunohistochemical characterization of neoplastic tissue, only pan-cytokeratin proved cross-reactive. Electron microscopy was performed in four cases. Tumours should be considered as differential diagnosis in cases with buoyancy issues, debilitated or emaciated animals, and may predispose to secondary infections. This study highlights the value of histopathological disease surveillance for commercial aquarium settings.

A variety of imaging and analytical methods have been developed to study nanoparticles in cells. Each has its benefits, limitations, and varying degrees of expense and difficulties in implementation. High-resolution analytical scanning transmission electron microscopy (HRSTEM) has the unique ability to image local cellular environments adjacent to a nanoparticle at near atomic resolution and apply analytical tools to these environments such as energy dispersive spectroscopy and electron energy loss spectroscopy. These tools can be used to analyze particle location, translocation and potential reformation, ion dispersion, and in vivo synthesis of second-generation nanoparticles. Such analyses can provide in depth understanding of tissue-particle interactions and effects that are caused by the environmental \"invader\" nanoparticles. Analytical imaging can also distinguish phases that form due to the transformation of \"invader\" nanoparticles in contrast to those that are triggered by a response mechanism, including the commonly observed iron biomineralization in the form of ferritin nanoparticles. The analyses can distinguish ion species, crystal phases, and valence of parent nanoparticles and reformed or in vivo synthesized phases throughout the tissue. This article will briefly review the plethora of methods that have been developed over the last 20 years with an emphasis on the state-of-the-art techniques used to image and analyze nanoparticles in cells and highlight the sample preparation necessary for biological thin section observation in a HRSTEM. Specific applications that provide visual and chemical mapping of the local cellular environments surrounding parent nanoparticles and second-generation phases are demonstrated, which will help to identify novel nanoparticle-produced adverse effects and their associated mechanisms.

The morphological characteristics of tendons have been thoroughly evaluated via microscopy. Optical microscopy and electron microscopy are the most commonly used techniques for tendon tissue observation. According to the principles of both microscopy types, preparation and evaluation methods vary. Simple optical microscopy is commonly used in the observation of cells and extracellular matrix, and many stains, including hematoxylin-eosin, Van Gieson, Prussian blue, Alcian blue, and toluidine blue, are used for evaluating cells, collagen fiber arrangement, and noncollagenous proteins. Histological scoring systems have been used in many studies for semi-quantification. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) are the most commonly used electron microscopy types, and special consideration is needed for the fixation and embedding protocols. Glutaraldehyde followed by osmium is most commonly used in the chemical fixation of tendon tissue, followed by epoxy resin embedment. Longitudinal sections captured in SEM images show the arrangement of collagen fibrils and the cells and lipid drops among them, while cross sections captured in TEM images show the diameter and distribution of collagen fibrils. SEM and TEM are used together for comprehensive evaluations. This mini review is focused on the preparation methodology and related evaluation indexes for the morphological evaluation of tendons.

High-resolution transmission electron microscopy (HRTEM) is an important approach to analyzing material structures. However, in reality, preparing a sufficiently thin sample for use in HRTEM, based on which images could be interpreted by weak phase object approximation theory, is difficult. During the imaging process, the thickness of the sample has two primary effects-a dynamical effect and a non-linear effect. Both are reviewed in this paper. Considering only the dynamical effect, the Bloch wave method and multislice theory have been proposed to understand the relationship between sample thickness and imaging. These methods exhibit high accuracy but high complexity as well. Sacrificing accuracy, pseudo-weak phase object approximation (PWPOA) theory can provide clues to the relationship in reciprocal space with greater simplicity. Meanwhile, in real space, channeling theory describes the dynamical effect with sufficient accuracy, and with the 1s state approximation, i.e., for a certain range of thicknesses, it provides a physical image and simplified expression with which to describe the relationship between the exit wave and sample thickness. As for the non-linear effect, a method of separating linear and non-linear information using a combination of transmission cross-coefficient theory and PWPOA theory was recently proposed. The variation of non-linear and linear imaging with sample thickness has also been discussed. A deep understanding has been acquired regarding the effects of the sample thickness, but a complete understanding of the HRTEM imaging process for thick samples has remained elusive. This understanding is crucial to the retrieval of structure from HRTEM images.

Protecting resin-dentin interfaces from hydrolytic and enzymatic degradation is critical for the longevity of adhesive restorations. In recent years, several strategies have been tested in vitro to induce apatite precipitation within interfibrillar and intrafibrillar collagen spaces, as well as in resin-sparse regions where the adhesive infiltration was incomplete. Also, the presence of calcium ions and other metallic ions has shown an inhibitory effect on enzymatic activity. Ion-releasing particles and biomimetic analogs have been studied for hybrid layer remineralization. Overall, remineralization strategy is dependent on the remaining mineral content. In partially demineralized dentin, residual apatite crystallites serve as nucleation sites for calcium and phosphate ions precipitation and crystal growth (\"top-down\" remineralization). In completely demineralized dentin where crystallites are absent (e.g., acid etched dentin) the use of mineral nano-precursors assisted by non-collagenous proteins analogs are necessary (\"bottom-up\" remineralization). This article reviews the approaches for hybrid layer remineralization and resin-dentin interface preservation.