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Resolution enhancement of total internal reflection microscopy via polarised scattering.
IF 1.5 4区 工程技术
Journal of microscopy Pub Date : 2025-03-05 DOI: 10.1111/jmi.13396
Chenguang Liu, Zhao Chen, Zhenlong Xu, Qinhai Yang, Jian Liu
{"title":"Resolution enhancement of total internal reflection microscopy via polarised scattering.","authors":"Chenguang Liu, Zhao Chen, Zhenlong Xu, Qinhai Yang, Jian Liu","doi":"10.1111/jmi.13396","DOIUrl":"https://doi.org/10.1111/jmi.13396","url":null,"abstract":"<p><p>This paper presents a novel approach for enhancing the resolution of label-free total internal reflection microscopy. This method leverages the directional difference of the sample polar scattering by capturing microscopic images under the illumination of evanescent waves propagating in various directions. By computing the high-order autocorrelation accumulation of these images at different angles, we achieved a 1.5-fold improvement in resolution while effectively reducing the distortion caused by laser speckles and image artefacts. This technique holds promise for long-term, noninvasive observation with enhanced resolution.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-03-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143557196","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}
引用次数: 0
Spatiotemporal visualisation of electrocatalyst/electrolyte interfaces with electrochemical atomic force microscopy: Applications and notes.
IF 1.5 4区 工程技术
Journal of microscopy Pub Date : 2025-03-03 DOI: 10.1111/jmi.13401
Weiran Zheng
{"title":"Spatiotemporal visualisation of electrocatalyst/electrolyte interfaces with electrochemical atomic force microscopy: Applications and notes.","authors":"Weiran Zheng","doi":"10.1111/jmi.13401","DOIUrl":"https://doi.org/10.1111/jmi.13401","url":null,"abstract":"<p><p>Electrochemical Atomic Force Microscopy (EC-AFM) has become a powerful tool for visualising dynamic processes at electrode/electrolyte interfaces with a nanoscale resolution. This technique enables real-time monitoring of morphological, chemical, and structural changes in electrocatalysts under operating conditions, providing critical insights into the mechanisms of electrocatalytic reactions. In this review, I introduce some applications of EC-AFM in electrocatalysis research and experimental considerations. The applications include tracking catalyst surface reconstruction, adsorption/desorption dynamics of intermediate species, long-range probing of the electrochemical interface, and catalyst degradation analysis. Moreover, experimental challenges, including cantilever selection, liquid-phase imaging stability, and artefacts, are discussed. Bridging the gap between nanoscale imaging and electrochemical analysis, EC-AFM offers a unique pathway to unravel complex interfacial phenomena critical for the design of next-generation electrocatalysts.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143542370","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}
引用次数: 0
Research on the SEGDC-UNet electron microscope image segmentation algorithm based on channel attention mechanism.
IF 1.5 4区 工程技术
Journal of microscopy Pub Date : 2025-02-27 DOI: 10.1111/jmi.13394
Yue Li, Qian Zhao, Haijing Sun, Yichuan Shao, Yong Wang
{"title":"Research on the SEGDC-UNet electron microscope image segmentation algorithm based on channel attention mechanism.","authors":"Yue Li, Qian Zhao, Haijing Sun, Yichuan Shao, Yong Wang","doi":"10.1111/jmi.13394","DOIUrl":"https://doi.org/10.1111/jmi.13394","url":null,"abstract":"<p><p>In this paper, we propose SEGDC-UNet, a segmentation algorithm for electron microscope (EM) images based on the channel attention mechanism. This algorithm integrates the channel attention mechanism and the GELU activation function into the DC-UNet network. By leveraging global information to selectively enhance primary features and suppress less relevant ones, the channel attention mechanism improves focus on important image channels and characteristics. Additionally, the GELU activation function enhances training performance and convergence speed. To evaluate its effectiveness, we compar SEGDC-UNet with six major lightweight image segmentation models on EMPS-Augmented electron microscopy image dataset. Experimental results demonstrate that the SEGDC-UNet model achieves higher Dice coefficient, IoU, Pixel Accuracy and Recall in electron microscopy image segmentation.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143515867","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}
引用次数: 0
De-risking transformative microscopy technologies for broad adoption.
IF 1.5 4区 工程技术
Journal of microscopy Pub Date : 2025-02-27 DOI: 10.1111/jmi.13400
Jesse Aaron, Teng-Leong Chew
{"title":"De-risking transformative microscopy technologies for broad adoption.","authors":"Jesse Aaron, Teng-Leong Chew","doi":"10.1111/jmi.13400","DOIUrl":"https://doi.org/10.1111/jmi.13400","url":null,"abstract":"<p><p>The past 20 years have seen a paradigm-shifting explosion of new optical microscopy technologies aimed at uncovering fundamental biological insights. Yet only a small portion 'cross the finish line' into wide adoption by the life science community. We contend that this is not primarily due to a lack of technical prowess or utility. Rather, many risks can conspire to derail the adoption of potentially disruptive technologies. One way to address these challenges is to de-risk paradigm-shifting inventions within open-access technology incubators. Here we detail the framework needed to shepherd innovative microscopy techniques through the often-treacherous adoption landscape to enable transformative scientific output.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143515753","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}
引用次数: 0
AutoSAFT: Autofocusing for extended depth of imaging in scanning acoustic microscopy.
IF 1.5 4区 工程技术
Journal of microscopy Pub Date : 2025-02-25 DOI: 10.1111/jmi.13399
M Sarim Ameed Khan, Shivam Ojha, Shubham Kumar Gupta, Amit Shelke, Azeem Ahmad, Frank Melandsø, Anowarul Habib
{"title":"AutoSAFT: Autofocusing for extended depth of imaging in scanning acoustic microscopy.","authors":"M Sarim Ameed Khan, Shivam Ojha, Shubham Kumar Gupta, Amit Shelke, Azeem Ahmad, Frank Melandsø, Anowarul Habib","doi":"10.1111/jmi.13399","DOIUrl":"https://doi.org/10.1111/jmi.13399","url":null,"abstract":"<p><p>In the scanning acoustic microscopy (SAM) imaging, it is essential to address the diminished lateral resolution in the out-of-focus region, as image quality correlates with the ultrasound propagation distance either above or below the focal plane. To focus the scanned image, a refocusing technique called synthetic aperture focusing technique (SAFT) is widely used, which improves the resolution by extending the depth of focus manually. In SAM, refocusing the image accurately is challenging without prior defocusing information. This paper introduces AutoSAFT, an automated version of SAFT. It employs the reference-less image-quality index (IQI) called Blind/Referenceless Image Spatial Quality Evaluator (BRISQUE) score to evaluate and optimise the defocusing distance automatically. The focused images were qualitatively analysed using metrics such as structural similarity index matrix (SSIM) and peak signal-to-noise ratio (PSNR). The combined qualitative and quantitative analysis demonstrates that the AutoSAFT technique is the most suitable method for the automatic focusing of acoustic imaging from the SAM. The proposed AutoSAFT opens a new avenue in photoacoustic image focusing methodologies.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143502059","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}
引用次数: 0
A 3D-printed optical microscope for low-cost histological imaging.
IF 1.5 4区 工程技术
Journal of microscopy Pub Date : 2025-02-25 DOI: 10.1111/jmi.13398
Jay Christopher, Rebecca Craig, Rebecca E McHugh, Andrew J Roe, Ralf Bauer, Brian Patton, Gail McConnell, Liam M Rooney
{"title":"A 3D-printed optical microscope for low-cost histological imaging.","authors":"Jay Christopher, Rebecca Craig, Rebecca E McHugh, Andrew J Roe, Ralf Bauer, Brian Patton, Gail McConnell, Liam M Rooney","doi":"10.1111/jmi.13398","DOIUrl":"https://doi.org/10.1111/jmi.13398","url":null,"abstract":"<p><p>We present the manufacture and characterisation of a 3D-printed, low-cost optical microscope using both a 3D-printed chassis and 3D-printed illumination and imaging optics. The required commercial components, consisting of a basic camera for image acquisition and light emitting diode controlled by a Raspberry Pi for illumination, are integrated into the 3D-printed microscope with the full design shown for ease of replication. Our 3D-printed microscope uses a single 3D-printed objective lens with a 2.9× magnification and a numerical aperture of 0.07. To benchmark the imaging performance of the system, we used standard test targets and histological specimens, namely, a Giemsa-stained blood smear sample and a thin section of mouse kidney stained with haemotoxylin and eosin. We demonstrated that subcellular resolution was obtained, and we corroborated this by imaging individual red blood cells and intricate anatomical details of the stained mouse kidney section. All of this was achieved using entirely 3D-printed hardware and optics, at a fraction of the cost of a commercial bright-field microscope, while presenting remarkable potential for customisation and increased accessibility for diagnostic imaging applications.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143492417","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}
引用次数: 0
Quantitative analysis of human umbilical vein endothelial cell morphology and tubulogenesis.
IF 1.5 4区 工程技术
Journal of microscopy Pub Date : 2025-02-21 DOI: 10.1111/jmi.13397
Viviane Mignone, Maria Augusta Arruda, Laura Kilpatrick, Benjamin Moore, Jeanette Woolard, Stephen Hill, Joëlle Goulding
{"title":"Quantitative analysis of human umbilical vein endothelial cell morphology and tubulogenesis.","authors":"Viviane Mignone, Maria Augusta Arruda, Laura Kilpatrick, Benjamin Moore, Jeanette Woolard, Stephen Hill, Joëlle Goulding","doi":"10.1111/jmi.13397","DOIUrl":"https://doi.org/10.1111/jmi.13397","url":null,"abstract":"<p><p>Primary human umbilical vein endothelial cells can grow as both a monolayer in culture and also as a capillary-like network making them an ideal model system in order to study vascular remodelling. Image-based analysis can allow assessment of cell morphology and motility but is dependent on accurate cell segmentation which requires high-contrast images not normally achievable without fluorescent markers. Here, ptychography is employed as a label-free image-based modality in order to extract quantitative metrics of morphology and tubulogenesis from cultured HUVECs over time in an automated multiwell assay. Phase-specific parameters of dry mass, optical thickness and sphericity were extracted and assessed alongside other metrics of cell number and shape. Tubulogenesis could be captured dynamically without any imaging artefacts from use of a basement membrane matrix and metrics of tube number, growth and branching exported alongside morphology metrics at early time-points. Utilising ptychography-based image analysis, all VEGF<sub>165</sub>a isoforms studied, elicited a concentration-dependent effect on cell elongation and survival within a HUVEC monolayer. Pharmacologically relevant parameters of potency (EC<sub>50</sub>) and efficacy were derived, exemplifying this label-free approach for the multiparameter and multiwell quantitative study of vascular remodelling in physiologically relevant cells at 37°C.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143468341","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}
引用次数: 0
Uncertainties in interfacial excess calculations from atom probe tomography data. 从原子探针断层扫描数据计算界面过量的不确定性。
IF 1.5 4区 工程技术
Journal of microscopy Pub Date : 2025-02-21 DOI: 10.1111/jmi.13392
Levi Tegg, Julie M Cairney
{"title":"Uncertainties in interfacial excess calculations from atom probe tomography data.","authors":"Levi Tegg, Julie M Cairney","doi":"10.1111/jmi.13392","DOIUrl":"https://doi.org/10.1111/jmi.13392","url":null,"abstract":"<p><p>Atom probe tomography can be used to measure the excess of solute chemical species at internal interfaces, but common protocols do not usually consider the compositional uncertainty of the (usually dilute) solute. Here, general models are derived for the composition profile of a solute segregated to the interface between two grains or phases. This model is fit to experimental data, with the cumulative compositional uncertainty included in the calculation of the interfacial excess. It is shown that the relative uncertainty in the interfacial excess can be large for dilute solutes, even when segregation is obvious in the concentration profile. Different methods for estimating the extent of the boundary provide different estimates for the interfacial excess and its uncertainty. Some strategies are provided for the minimisation of uncertainties in typical data, though ultimately, a typical calculation will be limited by compositional uncertainty arising from the relatively few solute counts in the region-of-interest.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143468403","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}
引用次数: 0
Modulation of host cell membrane biophysics dynamics by Neospora caninum: A study using LAURDAN fluorescence with hyperspectral imaging and phasor analysis.
IF 1.5 4区 工程技术
Journal of microscopy Pub Date : 2025-02-20 DOI: 10.1111/jmi.13395
Marcela Díaz, Carlos Robello, Andrés Cabrera, Leonel Malacrida
{"title":"Modulation of host cell membrane biophysics dynamics by Neospora caninum: A study using LAURDAN fluorescence with hyperspectral imaging and phasor analysis.","authors":"Marcela Díaz, Carlos Robello, Andrés Cabrera, Leonel Malacrida","doi":"10.1111/jmi.13395","DOIUrl":"https://doi.org/10.1111/jmi.13395","url":null,"abstract":"<p><p>Neospora caninum is known to manipulate host cell organelles and recruit lipids for its survival. However, the impact of this lipid redistribution on host cell membranes remains poorly understood. This study used LAURDAN fluorescence, hyperspectral imaging, and phasor plot analysis to investigate how N. caninum modifies membrane order in Vero cells. The results revealed a significant decrease in host cell plasma and internal membrane order upon infection, suggesting that cholesterol is redistributed from the host plasma membrane to the parasitophorous vacuoles. To mimic cholesterol depletion, uninfected cells were treated with methyl-β-cyclodextrin (MBCD), which increased membrane fluidity. Conversely, replenishing infected cells with cholesterol-loaded MBCD restored membrane fluidity to levels lower than control cells, indicating cholesterol enrichment. These findings provide novel insights into how N. caninum modulates host cell membrane dynamics through lipid manipulation, potentially aiding its intracellular survival.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143457835","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}
引用次数: 0
Investigation of sedimentary rocks and grains in the round: Micron-scale SEM-BEX 360° and 3D imaging.
IF 1.5 4区 工程技术
Journal of microscopy Pub Date : 2025-02-11 DOI: 10.1111/jmi.13389
Jim Buckman
{"title":"Investigation of sedimentary rocks and grains in the round: Micron-scale SEM-BEX 360° and 3D imaging.","authors":"Jim Buckman","doi":"10.1111/jmi.13389","DOIUrl":"https://doi.org/10.1111/jmi.13389","url":null,"abstract":"<p><p>The aim of the current paper is to demonstrate the use of a novel prototype horizontal rotation stage, within a scanning electron microscope (SEM). Here we utilise sedimentary rock chips and grains, with images collected every 10 ° of rotation, to produce a continuous overlapping field of view 360° montage. The same procedure can also be used to collect elemental X-ray maps. The collected images and maps can be simply combined into an image stack and saved as a movie format of the rotating specimen through 360°. More significantly, individual images and elemental maps can be reconstructed using photogrammetry imaging software, producing three-dimensional (3D) models with mesh worked surfaces and rendered texture (colour or greyscale) based on SEM image and X-ray data. This produces photorealistic smoothly transitioning movies with 360° fields of view around each object and can also be saved for stereo-viewing using a virtual reality (VR) headset. This new technique generates highly detailed information on the 3D structural and compositional relationships between components at the submicron to millimetre scale. The technique has widespread applications, including imaging other geological materials, biological mineralised tests and material science specimens such as metals, ceramics and building materials.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143399324","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}
引用次数: 0
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