Applied Microscopy

Abstract Glass vials are the most widely used primary containers for the packaging of parenteral products due to their optical clarity, general inertness, and hermetic properties, but under certain circumstances, they can pose safety concerns. Most of these issues are related to the potential formation of glass particulates through delamination or precipitation, resulting from the chemical interaction between the drug product and the inner surface of the glass vial. Hence, it is imperative for pharmaceutical companies to conduct product-vial compatibility studies to determine the appropriate packaging/container closure system. To support this development activity, scientists need to develop analytical methods to detect subvisible glass particulates in parenteral products, along with the appropriate positive controls, to facilitate detection and identification. This paper outlines the utilization of coaxial/episcopic and oblique illumination microscopy, combined with spectroscopic techniques, to detect thin glass particulates generated from a modified procedure. It also showcases the importance of angle-dependent lighting in visualizing positive control samples containing thin glass particulates. The analytical microscopy techniques discussed in this paper can assist scientists in selecting suitable container closure systems for developing parenteral products.

Adedayo M Sanni

Adedamola A Opalade

Armen Shamirian

Spencer Mattson

Eric Driscoll

Michael St Martin

Shikhar Mohan

Brooke Trimmer

Tarq Bunch

Robert Ovadia

Jungjoo Yoon

Sarina Ma

Chris Foti

Han-guk hyeonmigyeong hakoeji/Applied microscopy

Analytical microscopy techniques using coaxial and oblique illuminations to detect thin glass particulates generated from glass vials for parenteral drug products

用于检测肠外药物产品玻璃瓶产生的薄玻璃颗粒的同轴和斜照明分析显微技术

Abstract We examined the morphology of fertilized egg and ultrastructures of fertilized egg envelopes of dwarf rainbowfish ( Melanotaenia praecox ) belong to Melanotaeniidae using light and electron microscopes. The fertilized eggs were spherical with adhesive filament, transparent, demersal, and had a narrow perivitelline space and small oil droplets. The size of fertilized egg was 1.02 ± 0.18 mm ( n = 30), and there were two kinds of adhesive filament on the fertilized eggs. The long and thick (diameter 12.22 ± 0.52 μm, n = 20) adhesive filaments were only at the area of animal pole, and short and thin (diameter 1.99 ± 0.23 μm, n = 20) adhesive filaments were around the long filaments. A micropyle was conical shaped with adhesive filament and located near the animal pole of egg. The outer surface of fertilized egg was rough side. Also, the total thickness of the fertilized egg envelope was about 7.46 ± 0.41 μm ( n = 20), the fertilized egg envelope consisted of two layers, an inner lamellae layer and an outer layer with high electron-density. And the inner layer was 8 layers. Collectively, these morphological characteristics and adhesive property of fertilized egg with adhesive filaments, and ultrastructures of micropyle, outer surface, and section of fertilized egg envelope are showed species specificity.

Joon Hyung Sohn

Dong Heui Kim

Ultrastructure of the fertilized egg envelope from Melanotaenia praecox, Melanotaeniidae, Teleostei

条鳉科鱼类梅氏条鳉受精卵卵膜的超微结构

Abstract The olfactory organs of two African lungfishes, Protopterus amphibius and P. dolloi , were investigated using a stereo microscope and a compound light microscope and were described anatomically, histologically, and histochemically. Like other lungfishes, these species present the following general features: i) elongated olfactory chamber (OC), ii) anterior nostril at the ventral tip of the upper lip, iii) posterior nostril on the palate of the oral cavity, iv) lamellae with multiple cell types such as olfactory receptor neurons, supporting cells, basal cells, lymphatic cells, and mucous cells (MC), and vi) vomero-like epithelial crypt (VEC) made of glandular epithelium (GE) and crypt sensory epithelium. Some of these features exhibit differences between species: MCs are abundant in both the lamellar and inner walls of the OC in P. amphibius but occur only in lamellae in P. dolloi . On the other hand, some between feature differences are consistent across species: the GE of both P. amphibius and P. dolloi is strongly positive for Alcian blue (pH 2.5)-periodic acid Schiff (deep violet coloration), and positive with hematoxylin and eosin and with Masson’s trichrome (reddish-brown staining), unlike the MCs of the two species which stain dark red with both Alcian blue (pH 2.5)-periodic acid Schiff and Masson’s trichrome but respond faintly to hematoxylin and eosin. The differing abundance of MCs in the two lungfishes might reflect different degrees in aerial exposure of the olfactory organ, while the neutral and acid mucopolysaccharide-containing VEC, as indicated by staining properties of the MCs, is evolutionary evidence that P. amphibius and P. dolloi are the closest living relatives to tetrapods, at least in the order Dipnoi.

Hyun Tae Kim

Jong Young Park

Morphology and histology of the olfactory organ of two African lungfishes, Protopterus amphibius and P. dolloi (Lepidosirenidae, Dipnoi)

非洲肺鱼（Protopterus amphibius 和 P. dolloi）嗅觉器官的形态学和组织学研究（Lepidosirenidae, Dipnoi）

Abstract Histopathology is a well-established standard diagnosis employed for the majority of malignancies, including breast cancer. Nevertheless, despite training and standardization, it is considered operator-dependent and errors are still a concern. Fractal dimension analysis is a computational image processing technique that allows assessing the degree of complexity in patterns. We aimed here at providing a robust and easily attainable method for introducing computer-assisted techniques to histopathology laboratories. Slides from two databases were used: A) Breast Cancer Histopathological; and B) Grand Challenge on Breast Cancer Histology. Set A contained 2480 images from 24 patients with benign alterations, and 5429 images from 58 patients with breast cancer. Set B comprised 100 images of each type: normal tissue, benign alterations, in situ carcinoma, and invasive carcinoma. All images were analyzed with the FracLac algorithm in the ImageJ computational environment to yield the box count fractal dimension (Db) results. Images on set A on 40x magnification were statistically different ( p = 0.0003), whereas images on 400x did not present differences in their means. On set B, the mean Db values presented promissing statistical differences when comparing. Normal and/or benign images to in situ and/or invasive carcinoma (all p &lt; 0.0001). Interestingly, there was no difference when comparing normal tissue to benign alterations. These data corroborate with previous work in which fractal analysis allowed differentiating malignancies. Computer-aided diagnosis algorithms may beneficiate from using Db data; specific Db cut-off values may yield ~ 99% specificity in diagnosing breast cancer. Furthermore, the fact that it allows assessing tissue complexity, this tool may be used to understand the progression of the histological alterations in cancer.

Lucas Glaucio da Silva

Waleska Rayanne Sizinia da Silva Monteiro

Tiago Medeiros de Aguiar Moreira

Maria Aparecida Esteves Rabelo

Emílio Augusto Campos Pereira de Assis

Gustavo Torres de Souza

Fractal dimension analysis as an easy computational approach to improve breast cancer histopathological diagnosis

分形维数分析作为一种简便的计算方法以改进乳腺癌组织病理学诊断

Abstract Neuromorphic systems require integrated structures with high-density memory and selector devices to avoid interference and recognition errors between neighboring memory cells. To improve the performance of a selector device, it is important to understand the characteristics of the switching process. As changes by switching cycle occur at local nanoscale areas, a high-resolution analysis method is needed to investigate this phenomenon. Atomic force microscopy (AFM) is used to analyze the local changes because it offers nanoscale detection with high-resolution capabilities. This review introduces various types of AFM such as conductive AFM (C-AFM), electrostatic force microscopy (EFM), and Kelvin probe force microscopy (KPFM) to study switching behaviors.

Young-Min Kim

Jihye Lee

Deok-Jin Jeon

Si-Eun Oh

Jong-Souk Yeo

Advanced atomic force microscopy-based techniques for nanoscale characterization of switching devices for emerging neuromorphic applications

基于原子力显微镜的高级技术在新型神经形态应用开关器件纳米尺度表征中的应用

Muhammad Ishtiaq

Aqil Inam

Saurabh Tiwari

Jae Bok Seol

Appl Microsc

Correction: Microstructural, mechanical, and electrochemical analysis of carbon doped AISI carbon steels

更正：掺碳AISI碳钢的微观结构、力学性能和电化学分析

Oxide-supported metal catalysts are essential components in industrial processes for catalytic conversion. However, the performance of these catalysts is often compromised in high temperature reaction environments due to sintering effects. Currently, a number of studies are underway with the objective of improving the metal support interaction (MSI) effect in order to enhance sintering resistance by surface modification of the oxide support, including the formation of inhomogeneous defects on the oxide support, the addition of a rare earth element, the use of different facets, encapsulation, and other techniques. The recent developments in in situ gas phase transmission electron microscopy (TEM) have enabled direct observation of the sintering process of NPs in real time. This capability further allows to verify the efficacy of the methods used to tailor the support surface and contributes effectively to improving sintering resistance. Here, we review a few selected studies on how in situ gas phase TEM has been used to prevent the sintering of catalyst NPs on oxide supports.

Wonjun Kim

Kangsik Kim

Jaejin Kim

Zonghoon Lee

In situ observation of catalyst nanoparticle sintering resistance on oxide supports via gas phase transmission electron microscopy

通过气相透射电子显微镜原位观察氧化物载体上催化剂纳米颗粒的烧结抗性

Abstract The sliding filament theory and the cross-bridge model have been fundamental in understanding muscle contraction. While the cross-bridge model explains the interaction between a single myosin head and actin filament, the native myosin molecule consists of two heads. This review explores the possibility and mechanism of two-headed binding in myosin II to the actin. Recent studies using electron tomography and resonance energy transfer have provided evidence in support of the occurrence of two-headed binding. The flexibility of the regulatory light chain (RLC) appears to play a significant role in enabling this binding mode. However, high-resolution structures of the RLCs in the two-headed bound state have not yet been reported. Resolving these structures, possibly through sub-tomogram averaging or single-particle analysis, would provide definitive proof of the conformational flexibility of RLCs and their role in facilitating two-headed binding. Further investigations are also required to address questions such as the predominance of two-headed versus single-headed binding and the influence of the state of each of the heads on the other. An understanding of the mechanism of two-headed binding is crucial for developing a comprehensive model of the cross-bridge cycle of the native myosin molecule.

Yoon Ho Park

Gang San Song

Hyun Suk Jung

Research reviews on myosin head interactions with F-actin

肌球蛋白头部与F-肌动蛋白相互作用的研究综述

Plant cells are uniquely characterized by exhibiting cell walls, pigments, and phenolic compounds, which can impede microscopic observations by absorbing and scattering light. The concept of clearing was first proposed in the late nineteenth century to address this issue, aiming to render plant specimens transparent using chloral hydrate. Clearing techniques involve chemical procedures that render biological specimens transparent, enabling deep imaging without physical sectioning. Drawing inspiration from clearing techniques for animal specimens, various protocols have been adapted for plant research. These procedures include (i) hydrophobic methods (e.g., Visikol™), (ii) hydrophilic methods (ScaleP and ClearSee), and (iii) hydrogel-based methods (PEA-CLARITY). Initially, clearing techniques for plants were mainly utilized for deep imaging of seeds and leaves of herbaceous plants such as Arabidopsis thaliana and rice. Utilizing cell wall-specific fluorescent dyes for plants and fungi, researchers have documented the post-penetration behavior of plant pathogenic fungi within hosts. State-of-the-art plant clearing techniques, coupled with microbe-specific labeling and high-throughput imaging methods, offer the potential to advance the in planta characterization of plant microbiomes.

Ki Woo Kim

Clearing techniques for deeper imaging of plants and plant–microbe interactions

植物和植物-微生物相互作用深层成像的清除技术

ISSN	2234-6198, 2287-4445, 2287-5123
h-index	10

出版信息	出版商: Springer Nature，出版周期: ，期刊类型: journal，开源期刊: 否
基本数据	创刊年份: 2019，原创研究文献占比: ，自引率:， Gold OA占比:

Applied Microscopy

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Analytical microscopy techniques using coaxial and oblique illuminations to detect thin glass particulates generated from glass vials for parenteral drug products

Ultrastructure of the fertilized egg envelope from Melanotaenia praecox, Melanotaeniidae, Teleostei

Morphology and histology of the olfactory organ of two African lungfishes, Protopterus amphibius and P. dolloi (Lepidosirenidae, Dipnoi)

Fractal dimension analysis as an easy computational approach to improve breast cancer histopathological diagnosis

Advanced atomic force microscopy-based techniques for nanoscale characterization of switching devices for emerging neuromorphic applications

Analytical microscopy techniques using coaxial and oblique illuminations to detect thin glass particulates generated from glass vials for parenteral drug products

Correction: Microstructural, mechanical, and electrochemical analysis of carbon doped AISI carbon steels

In situ observation of catalyst nanoparticle sintering resistance on oxide supports via gas phase transmission electron microscopy

Research reviews on myosin head interactions with F-actin

Clearing techniques for deeper imaging of plants and plant–microbe interactions