Journal of microscopy最新文献

{"title":"Automated Euler number of the alveolar capillary network based on deep learning segmentation with verification by stereological methods.","authors":"Julia Schmidt, Jonas Labode, Christoph Wrede, Yannick Regin, Jaan Toelen, Christian Mühlfeld","doi":"10.1111/jmi.13390","DOIUrl":"https://doi.org/10.1111/jmi.13390","url":null,"abstract":"<p><p>Diseases like bronchopulmonary dysplasia (BPD) affect the development of the pulmonary vasculature, including the alveolar capillary network (ACN). Since pulmonary development is highly dependent on angiogenesis and microvascular maturation, ACN investigations are essential. Therefore, efficient methods are needed for quantitative comparative studies. Here, the suitability of deep learning (DL) for processing serial block-face scanning electron microscopic (SBF-SEM) data by generating ACN segmentations, 3D reconstructions and performing automated quantitative analyses based on them, was tested. Since previous studies revealed inefficient ACN segmentation as the limiting factor in the overall workflow, a 2D DL-based approach was used with existing data, aiming at the reduction of necessary manual interaction. Automated quantitative analyses based on completed segmentations were performed subsequently. The results were compared to stereological estimations, assessing segmentation quality and result reliability. It was shown that the DL-based approach was suitable for generating segmentations on SBF-SEM data. This approach generated more complete initial ACN segmentations than an established method, despite the limited amount of available training data and the use of a 2D rather than a 3D approach. The quality of the completed ACN segmentations was assessed as sufficient. Furthermore, quantitative analyses delivered reliable results about the ACN architecture, automatically obtained contrary to manual stereological approaches. This study demonstrated that ACN segmentation is still the part of the overall workflow that requires improvement regarding the reduction of manual interaction to benefit from available automated software tools. Nevertheless, the results indicated that it could be advantageous taking further efforts to implement a 3D DL-based segmentation approach. As the amount of analysed data was limited, this study was not conducted to obtain representative data about BPD-induced ACN alterations, but to highlight next steps towards a fully automated segmentation and evaluation workflow, enabling larger sample sizes and representative studies.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143066044","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Introduction to special issue '12th International Botanical Microscopy Meeting'.","authors":"Kim Findlay","doi":"10.1111/jmi.13388","DOIUrl":"https://doi.org/10.1111/jmi.13388","url":null,"abstract":"","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143052815","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"ApoNecV: A macro for cell death type differentiation.","authors":"Marketa Kolarikova, Barbora Hosikova, Jiri Tesarik, Katerina Langova, Hana Kolarova","doi":"10.1111/jmi.13386","DOIUrl":"https://doi.org/10.1111/jmi.13386","url":null,"abstract":"<p><p>The evaluation of large experimental datasets is a fundamental aspect of research in every scientific field. Streamlining this process can improve the reliability of results while making data analysis more efficient and faster to execute. In biomedical research it is often very important to determine the type of cell death after various treatments. Thus, differentiating between viable, apoptotic, and necrotic cells provide critical insights into the treatment efficacy, a key aspect in the field of drug development. Fluorescent microscopy is perceived as a widely used technique for cell metabolism assessment and can therefore be used to investigate treatment outcomes after staining samples with cell death detection kit. However, accurate evaluation of therapeutic results requires quantitative analysis, often necessitating extensive postprocessing of imaging data. In this study, we introduce a complementary tool designed as a macro for the Fiji platform, enabling the automated postprocessing of fluorescent microscopy images to accurately distinguish and quantify viable, apoptotic, and necrotic cells.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143033370","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ultrastructure expansion microscopy: Enlarging our perspective on apicomplexan cell division.","authors":"Sofía Horjales, Florencia Sena, María E Francia","doi":"10.1111/jmi.13387","DOIUrl":"https://doi.org/10.1111/jmi.13387","url":null,"abstract":"<p><p>Apicomplexans, a large phylum of protozoan intracellular parasites, well known for their ability to invade and proliferate within host cells, cause diseases with major health and economic impacts worldwide. These parasites are responsible for conditions such as malaria, cryptosporidiosis, and toxoplasmosis, which affect humans and other animals. Apicomplexans exhibit complex life cycles, marked by diverse modes of cell division, which are closely associated with their pathogenesis. All the unique structural and evolutionary characteristics of apicomplexan parasites, the biology underlying life stage transitions, and the singular mechanisms of cell division alongside their associated biomedical relevance have captured the attention of parasitologists of all times. Traditional light and electron microscopy have set the fundamental foundations of our understanding of these parasites, including the distinction among their modes of cell division. This has been more recently complemented by microscopy advances through the implementation of superresolution fluorescence microscopy, and variants of electron microscopy, such as cryo-EM and tomography, revealing intricate details of organelles and cell division. Ultrastructure Expansion Microscopy has emerged as a transformative, accessible approach that enhances resolution by physically expanding samples isometrically, allowing nanoscale visualisation on standard light microscopes. In this work, we review the most recent contributions of U-ExM and its recent improvements and innovations, in providing unprecedented insights into apicomplexan ultrastructure and its associated mechanisms, focusing particularly on cell division. We highlight the power of U-ExM in combination with protein-specific labelling, in aiding the visualisation of long oversighted organelles and detailed insights into the assembly of parasite-specific structures, such as the conoid in Plasmodia, and the apical-basal axis in Toxoplasma, respectively, during new parasite assembly. Altogether, the contributions of U-ExM reveal conserved and unique structural features across species while nearing super resolution. The development of these methodologies and their combination with different technologies are crucial for advancing our mechanistic understanding of apicomplexan biology, offering new perspectives that may facilitate novel therapeutic strategies against apicomplexan-caused diseases.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-01-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143033373","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Automatic visual detection of activated sludge microorganisms based on microscopic phase contrast image optimisation and deep learning.","authors":"Dan Liang, Yuming Yao, Minjie Ye, Qinze Luo, Jiale Chu","doi":"10.1111/jmi.13385","DOIUrl":"https://doi.org/10.1111/jmi.13385","url":null,"abstract":"<p><p>The types and quantities of microorganisms in activated sludge are directly related to the stability and efficiency of sewage treatment systems. This paper proposes a sludge microorganism detection method based on microscopic phase contrast image optimisation and deep learning. Firstly, a dataset containing eight types of microorganisms is constructed, and an augmentation strategy based on single and multisamples processing is designed to address the issues of sample deficiency and uneven distribution. Secondly, a phase contrast image quality optimisation algorithm based on fused variance is proposed, which can effectively improve the standard deviation, entropy, and detection performance. Thirdly, a lightweight YOLOv8n-SimAM model is designed, which introduces a SimAM attention module to suppress the complex background interference and enhance attentions to the target objects. The lightweight of the network is realised using a detection head based on multiscale information fusion convolutional module. In addition, a new loss function IW-IoU is proposed to improve the generalisation ability and overall performance. Comparative and ablative experiments are conducted, demonstrating the great application potential for rapid and accurate detection of microbial targets. Compared to the baseline model, the proposed method improves the detection accuracy by 12.35% and hastens the running speed by 37.9 frames per second while evidently reducing the model size.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143023903","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In situ quantification of ribosome number by electron tomography.","authors":"Mounir El Hankouri, Marco Nousch, Aayush Poddar, Thomas Müller-Reichert, Gunar Fabig","doi":"10.1111/jmi.13380","DOIUrl":"https://doi.org/10.1111/jmi.13380","url":null,"abstract":"<p><p>Ribosomes, discovered in 1955 by George Palade, were initially described as small cytoplasmic particles preferentially associated with the endoplasmic reticulum (ER). Over the years, extensive research has focused on both the structure and function of ribosomes. However, a fundamental question - how many ribosomes are present within whole cells - has remained largely unaddressed. In this study, we developed a microscopic method to quantify the total number of ribosomes in hTERT-RPE-1 cells and in nematode cells from various tissues of Caenorhabditis elegans hermaphrodites. Using electron tomography of high-pressure frozen, freeze-substituted and resin-embedded samples, we determined that the ribosome number in hTERT-RPE-1 cells is in the same order of magnitude as biochemical measurements obtained via RNA capillary electrophoresis. As expected, control worms exhibited a higher number of ribosomes compared to RNA polymerase I A subunit (RPOA-1)-depleted worms in two out of three analysed tissue types. Our imaging-based approach complements established biochemical methods by enabling direct quantification of ribosome numbers in specific samples. This method offers a powerful tool for advancing our understanding of ribosome localisation and distribution in cells and tissues across diverse model systems.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142983702","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cryo-SEM and large volume FIB-SEM of Arabidopsis cotyledons: Degradation of lipid bodies, biogenesis of glyoxysomes and reorganisation of organelles during germination.","authors":"Gerhard Wanner, Elizabeth Schroeder-Reiter, Farhah F Assaad","doi":"10.1111/jmi.13381","DOIUrl":"https://doi.org/10.1111/jmi.13381","url":null,"abstract":"<p><p>Until recently, the lack of three-dimensional visualisation of whole cells at the electron microscopic (EM) level has led to a significant gap in our understanding of the interaction of cellular organelles and their interconnection. This is particularly true with regard to the role of the endoplasmic reticulum (ER). In this study, we perform three-dimensional reconstructions of serial FIB/SEM stacks and anaglyphs derived from volume rendering, cryo-scanning electron microscopy (cryo-SEM) and state-of-the-art electron microscopy immobilisation and imaging techniques. The results show that glyoxysomes are formed de novo in large numbers and in characteristic clusters on the ER upon germination in mesophyll cells of Arabidopsis cotyledons. The degradation of lipid bodies during germination occurs not only via the ER, which enlarges by taking up polar lipids resulting from enzymatic degradation by lipases, but also via glyoxysomes, which engulf lipid bodies. Dictyosomal (Golgi-derived) vesicles, which fuse with glyoxysomes or their precursors, also appear to be involved in the differentiation of glyoxysomes from segments of the ER. The formation of the central vacuole is the result of the fusion of protein storage vacuoles (protein bodies), which become complex three-dimensional structures during germination. Our observations also suggest that the vacuole plays a role in the degradation of glyoxysomes. The evidence provided in three dimensions shows that the endoplasmic reticulum plays a central role in the biogenesis and degradation of lipid bodies, the ontogeny of glyoxysomes and the development of plastids in the mesophyll cells of Arabidopsis cotyledons.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142971283","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Compressive electron backscatter diffraction imaging.","authors":"Zoë Broad, Alex W Robinson, Jack Wells, Daniel Nicholls, Amirafshar Moshtaghpour, Angus I Kirkland, Nigel D Browning","doi":"10.1111/jmi.13379","DOIUrl":"https://doi.org/10.1111/jmi.13379","url":null,"abstract":"<p><p>Electron backscatter diffraction (EBSD) has developed over the last few decades into a valuable crystallographic characterisation method for a wide range of sample types. Despite these advances, issues such as the complexity of sample preparation, relatively slow acquisition, and damage in beam-sensitive samples, still limit the quantity and quality of interpretable data that can be obtained. To mitigate these issues, here we propose a method based on the subsampling of probe positions and subsequent reconstruction of an incomplete data set. The missing probe locations (or pixels in the image) are recovered via an inpainting process using a dictionary-learning based method called beta-process factor analysis (BPFA). To investigate the robustness of both our inpainting method and Hough-based indexing, we simulate subsampled and noisy EBSD data sets from a real fully sampled Ni-superalloy data set for different subsampling ratios of probe positions using both Gaussian and Poisson noise models. We find that zero solution pixel detection (inpainting un-indexed pixels) enables higher-quality reconstructions to be obtained. Numerical tests confirm high-quality reconstruction of band contrast and inverse pole figure maps from only 10% of the probe positions, with the potential to reduce this to 5% if only inverse pole figure maps are needed. These results show the potential application of this method in EBSD, allowing for faster analysis and extending the use of this technique to beam sensitive materials.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142965355","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Live cell imaging of plant infection provides new insight into the biology of pathogenesis by the rice blast fungus Magnaporthe oryzae.","authors":"Berlaine G Quime, Lauren S Ryder, Nicholas J Talbot","doi":"10.1111/jmi.13382","DOIUrl":"https://doi.org/10.1111/jmi.13382","url":null,"abstract":"<p><p>Magnaporthe oryzae is the causal agent of rice blast, one of the most serious diseases affecting rice cultivation around the world. During plant infection, M. oryzae forms a specialised infection structure called an appressorium. The appressorium forms in response to the hydrophobic leaf surface and relies on multiple signalling pathways, including a MAP kinase phosphorelay and cAMP-dependent signalling, integrated with cell cycle control and autophagic cell death of the conidium. Together, these pathways regulate appressorium morphogenesis.The appressorium generates enormous turgor, applied as mechanical force to breach the rice cuticle. Re-polarisation of the appressorium requires a turgor-dependent sensor kinase which senses when a critical threshold of turgor has been reached to initiate septin-dependent re-polarisation of the appressorium and plant infection. Invasive growth then requires differential expression and secretion of a large repertoire of effector proteins secreted by distinct secretory pathways depending on their destination, which is also governed by codon usage and tRNA thiolation. Cytoplasmic effectors require an unconventional Golgi-independent secretory pathway and evidence suggests that clathrin-mediated endocytosis is necessary for their delivery into plant cells. The blast fungus then develops a transpressorium, a specific invasion structure used to move from cell-to-cell using pit field sites containing plasmodesmata, to facilitate its spread in plant tissue. This is controlled by the same MAP kinase signalling pathway as appressorium development and requires septin-dependent hyphal constriction. Recent progress in understanding the mechanisms of rice infection by this devastating pathogen using live cell imaging procedures are presented.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142965485","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Imaging malaria parasites across scales and time.","authors":"Julien Guizetti","doi":"10.1111/jmi.13384","DOIUrl":"https://doi.org/10.1111/jmi.13384","url":null,"abstract":"<p><p>The idea that disease is caused at the cellular level is so fundamental to us that we might forget the critical role microscopy played in generating and developing this insight. Visually identifying diseased or infected cells lays the foundation for any effort to curb human pathology. Since the discovery of the Plasmodium-infected red blood cells, which cause malaria, microscopy has undergone an impressive development now literally resolving individual molecules. This review explores the expansive field of light microscopy, focusing on its application to malaria research. Imaging technologies have transformed our understanding of biological systems, yet navigating the complex and ever-growing landscape of techniques can be daunting. This review offers a guide for researchers, especially those working on malaria, by providing historical context as well as practical advice on selecting the right imaging approach. The review advocates an integrated methodology that prioritises the research question while considering key factors like sample preparation, fluorophore choice, imaging modality, and data analysis. In addition to presenting seminal studies and innovative applications of microscopy, the review highlights a broad range of topics, from traditional techniques like white light microscopy to advanced methods such as superresolution microscopy and time-lapse imaging. It addresses the emerging challenges of microscopy, including phototoxicity and trade-offs in resolution and speed, and offers insights into future technologies that might impact malaria research. This review offers a mix of historical perspective, technological progress, and practical guidance that appeal to novice and advanced microscopists alike. It aims to inspire malaria researchers to explore imaging techniques that could enrich their studies, thus advancing the field through enhanced visual exploration of the parasite across scales and time.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-01-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142921844","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}