Journal of microscopy最新文献

{"title":"The Origins of Ploem's Filter Cube: A Pandora's Box","authors":"Jack Gordon,&nbsp;Kaleah Balcomb,&nbsp;Vinod Maseedupally,&nbsp;Maitreyee Roy","doi":"10.1111/jmi.70137","DOIUrl":"10.1111/jmi.70137","url":null,"abstract":"<p>Ploem's filter cube, central to modern epifluorescence microscopy, integrates three matched optical components: an excitation filter, an emission filter, and a dichroic mirror. Together, these elements enable the separation of excitation and emission light paths, producing high-contrast fluorescence images. Despite their importance, the origins of these optical filters and the concept of a matched filter set remain unattributed. This review traces the development of these principles to discoveries made half a century before the fluorescence microscope. Through a narrative historical analysis, we identify George Stokes, Wilhelm Haidinger, and David Brewster as key contributors whose studies of light, colour, and polarisation established the foundations of fluorescence filtering. We also show that the matched filter concept can be dated to 173 years ago and that early dichroic films predated Ploem's cube by more than a century. By uncovering these origins, this work highlights how early optical discoveries shaped the design of modern fluorescence microscopy.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":"304 3","pages":"266-276"},"PeriodicalIF":1.9,"publicationDate":"2026-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/jmi.70137","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148308496","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Introduction to the Festschrift for Guy Christopher Cox","authors":"Louise Cole,&nbsp;Colin Sheppard","doi":"10.1111/jmi.70189","DOIUrl":"10.1111/jmi.70189","url":null,"abstract":"","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":"304 3","pages":"191-195"},"PeriodicalIF":1.9,"publicationDate":"2026-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149194413","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":"Confocal and multiphoton microscopy: A selective review","authors":"Colin J. R. Sheppard","doi":"10.1111/jmi.70080","DOIUrl":"10.1111/jmi.70080","url":null,"abstract":"<p>The development of confocal microscopy and multiphoton microscopy from the 1970s to the 1990s is reviewed. This is the period when the modern form of these microscopes developed and became commercialised. Key topics include point versus line scanning, nonlinear microscopy, and the emergence of image scanning microscopy. Guy Cox's role in the development is outlined.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":"304 3","pages":"209-220"},"PeriodicalIF":1.9,"publicationDate":"2026-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147593010","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":"When biology meets materials science – Interdisciplinary applications of electron microscopy","authors":"Martin Saunders,&nbsp;Peta L. Clode,&nbsp;Enoch V. S. Wong,&nbsp;Anna-Lee Jessop,&nbsp;Gerd E. Shröder-Turk,&nbsp;Felix de Tombeur","doi":"10.1111/jmi.70086","DOIUrl":"10.1111/jmi.70086","url":null,"abstract":"<p>Research at the interface between biology and materials science creates challenges for electron microscopists. Everything from the sample preparation to the choice of imaging and analytical techniques and the interpretation of the resulting data refuses to sit comfortably within the domain of one discipline or the other. Ultimately, these problems are best addressed by an interdisciplinary team that brings together expertise from both sides. While working in this environment can be challenging, with the need to learn new approaches, new language, and step outsides one's comfort zone, the benefits can be significant. Here we highlight those benefits using three examples of our own interdisciplinary collaborations (involving nanoparticles and other inorganic materials in plants, and nanostructures in butterflies) to encourage more electron microscopy researchers to reach out and embrace their colleagues on the other side of the interdisciplinary divide.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":"304 3","pages":"221-232"},"PeriodicalIF":1.9,"publicationDate":"2026-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/jmi.70086","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147623191","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Microscopy insights into reef coral fluorescent proteins: Diversity, light energy transfer and biological function","authors":"Anya Salih","doi":"10.1111/jmi.70186","DOIUrl":"10.1111/jmi.70186","url":null,"abstract":"<p>Fluorescent proteins (FPs) and chromoproteins (CPs) of reef-building corals, which generate much of their vivid colouration, occupy a distinctive position at the interface of natural photonics, symbiosis biology and fluorescence microscopy. They include cyan, green, yellow-orange and red, photoconvertible, photoswitchable FPs and weakly emissive CP classes and are often expressed at unusually high local concentrations within patterned tissues. Coral tissues, therefore, provide a naturally evolved system in which fluorescence, scattering, re-absorption and proximity-dependent energy transfer can be examined in vivo. This review focuses on the light-driven functions of coral GFP-type proteins in symbiosis, treating them as host-controlled optical elements that shape the photon environment experienced by <i>Symbiodiniaceae</i>. Evidence is synthesised for two complementary outcomes: photoprotection under high irradiance and redistribution, wavelength transformation or spectral enhancement of light in low-light tissues and habitats. These functions are particularly relevant during thermal stress and bleaching, when loss of symbiont pigmentation alters the internal light field and can change host FP/CP expression; in several coral taxa, high host fluorescence has also been associated with reduced bleaching susceptibility. In corals, strong spectral overlap, oligomerisation, photoconversion, chromoprotein- dark states, tissue and skeletal scattering make interpretation based on steady-state fluorescence intensity alone unreliable. Microscopy was critical for distinguishing the spatial and photophysical mechanisms underlying these optical effects. Confocal microspectroscopy localised spectrally distinct GFP-type proteins within intact tissues and revealed the spatial organisation required for light-transfer phenomena. Fluorescence lifetime imaging microscopy (FLIM) provided a means of distinguishing radiative re-emission/reabsorption from proximity-dependent non-radiative transfer through changes in donor fluorescence lifetime. Building on this foundation, the review advances an expanded interpretation of coral FP function in which dense pigment ensembles may redistribute or dissipate excitation through a hierarchy of processes, from tissue-scale radiative transfer to nanometre-scale FRET and, potentially, ultrafast proton-coupled excited-state dynamics and excitation migration. This Festschrift review honors Associate Professor Guy Cox, whose contributions to quantitative fluorescence microscopy, fluorescence lifetime imaging and collaborative coral FP research were foundational to this field.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":"304 3","pages":"301-344"},"PeriodicalIF":1.9,"publicationDate":"2026-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/jmi.70186","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149298096","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Near-infrared confocal imaging reveals the spatial organisation of far-red photosynthesis in Shark Bay microbial mats and Halomicronema sp.","authors":"Anya Salih,&nbsp;Katherina Petrou,&nbsp;Michael Johnson,&nbsp;Alexander M. Macmillan,&nbsp;Gareth Kindler,&nbsp;Brendan P. Burns,&nbsp;Unnikrishnan Kuzhiumparambil,&nbsp;Anthony W. D. Larkum","doi":"10.1111/jmi.70174","DOIUrl":"10.1111/jmi.70174","url":null,"abstract":"<p>Far-red light (FRL) cyanobacteria occupy spectrally filtered habitats where visible wavelengths are depleted but far-red radiation persists. Shark Bay (Western Australia) stromatolitic microbial mats provide such a habitat, containing subsurface cyanobacterial communities in which chlorophyll <i>d/f</i> (Chl <i>d/f</i>)-containing phototrophs coexist with filamentous <i>Halomicronema</i>-like taxa adapted to far-red photosynthesis. In this <i>Journal of Microscopy</i> issue honouring Associate Professor Guy C. Cox, we used FRL-sensitive confocal imaging to visualise far-red cyanobacteria in intact Shark Bay mat samples, and combined microspectroscopy, lifetime microscopy, HPLC pigment analysis and synchrotron FTIR microspectroscopy to examine pigment organisation and FRL acclimation in cultured <i>Halomicronema</i> sp. ‘Clifton’. Confocal imaging with near-infrared-sensitive detection revealed abundant far-red fluorescent cyanobacteria within subsurface mat layers and enabled direct visualisation of Chl <i>d/f</i>-containing phototrophs in situ. In cultured <i>Halomicronema</i>, HPLC confirmed that Chl <i>f</i> accumulation was induced by FRL acclimation and absent under white light, validating assignment of the dominant 730–745 nm emission in FRL-grown cells to Chl <i>f</i>-enriched photosystems. Intracellular excitation–emission mapping showed that FRL acclimation did not produce complete pigment segregation, but generated a pronounced intracellular pigment gradient, with Chl <i>f</i>-associated fluorescence enriched in mid-cell regions and phycobiliprotein- and chlorophyll <i>a</i> (Chl <i>a</i>)-associated fluorescence enriched at cell poles. FLIM and phasor analyses showed that this spatial reorganisation was accompanied by altered excited-state behaviour, including strongest lifetime shortening in Chl <i>a</i>-associated channel, additional shortening in the far-red chlorophyll-enriched channel and increased heterogeneity of PC/APC-associated fluorescence. The absence of a distinct Chl <i>f</i>-only lifetime population indicates that Chl <i>f</i> remains functionally integrated within coupled photosynthetic complexes rather than acting as an isolated fluorescent sink. By linking optical stratification and far-red cyanobacterial diversity within Shark Bay microbial mats to pigment organisation and fluorescence dynamics in living <i>Halomicronema</i> cells, this study extends Guy Cox's microscopy-based exploration of photosynthetic microorganisms and demonstrates the power of modern spectral confocal microscopy for investigating photosynthesis at the far-red edge of oxygenic life.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":"304 3","pages":"277-300"},"PeriodicalIF":1.9,"publicationDate":"2026-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/jmi.70174","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149081077","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Advancing collagen-related pathology assessment through second-harmonic generation imaging","authors":"Jackson Rodrigues,&nbsp;Manikanth Karnati,&nbsp;Gagan Raju,&nbsp;Edwin Pious,&nbsp;Ankur Gogoi,&nbsp;Guan-Yu Zhuo,&nbsp;Nirmal Mazumder,&nbsp;Fu-Jen Kao","doi":"10.1111/jmi.70098","DOIUrl":"10.1111/jmi.70098","url":null,"abstract":"<p>Collagen remodelling and dysregulation are the hallmarks of diverse pathological conditions. In this context, second-harmonic generation (SHG) imaging has emerged as a powerful label-free modality for assessing collagen. This offers submicron resolution, intrinsic optical sectioning, and deeper imaging capabilities without the need for exogenous agents. Additionally, polarisation-resolved SHG (P-SHG) further enhances orientation-sensitive information on collagen architecture. This enables the elucidation of subtle structural alterations that are often invisible to conventional imaging techniques. This nonlinear technique has demonstrated utility in diagnosing diseases, particularly in assessing tumour progression, where collagen remodelling is correlated with disease severity and prognosis. Furthermore, quantitative image analysis, using metrics such as fibre density, orientation, anisotropy, and alignment, provides objective measures of collagen remodelling. P-SHG complements these analyses by enabling the retrieval of molecular susceptibility ratios (e.g., χ33/χ31), degrees of polarisation (DOLP), and orientation distribution functions, yielding more profound insights into the fibrillar ultrastructure and molecular organisation. These quantitative descriptors are being increasingly integrated into clinically relevant feature extraction methods. The incorporation of these features in artificial intelligence (AI) methods has enhanced SHG-based pathological assessment. AI-driven models can automatically classify tissues, detect pathological patterns, and correlate the collagen microstructure with clinical outcomes, addressing the bottleneck of manual interpretation and enhancing reproducibility. Future perspectives highlight the integration of SHG and P-SHG with deep-tissue multiphoton imaging, accompanied by explainable AI/ML-driven quantification, which will improve diagnostic precision, reproducibility, and clinical adoption. Ultimately, the convergence of optical innovation, deep learning, and translational engineering will lead to the establishment of collagen pathology assessment through SHG and P-SHG as reliable, accessible tools for diverse clinical applications.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":"304 3","pages":"233-247"},"PeriodicalIF":1.9,"publicationDate":"2026-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/jmi.70098","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147773926","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Using confocal microscopy and image processing to measure properties of sub-resolution cell structures: Application to cardiac transverse tubules","authors":"Mark B. Cannell,&nbsp;Cherrie H. T. Kong","doi":"10.1111/jmi.70120","DOIUrl":"10.1111/jmi.70120","url":null,"abstract":"<p>Confocal microscopes increase the resolution of images by rejecting unwanted data from the out of focus regions of the specimen. This incurs a penalty in the recorded signal to noise ratio that may be partially ameliorated by inputting additional information during image processing. This is commonly seen in digital deconvolution techniques, where the point spread function (PSF) of the microscope is used to refine the image. By accounting for the interaction between the microscope PSF and labelled objects, information about sub-resolution structures can be quantified. In the case of cardiac t-tubules, we show the detailed 3D organisation and estimates of the t-tubule diameter and approximately 50% of t-tubules are below the classical optical resolution limit. By obtaining local t-tubule volume (<i>V</i>) and surface area (<i>SA</i>) by using dual labelling, together with control of the PSF, a simple relationship between t-tubule radius and the <i>V</i>:<i>SA</i> ratio can be derived. Solute movement can also be analysed by accounting for the shape of the illumination field during fluorescence recovery after photobleaching (FRAP). Using these methods, we show that the properties of murine and lagomorph ventricular t-tubules are quite different.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":"304 3","pages":"248-256"},"PeriodicalIF":1.9,"publicationDate":"2026-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/jmi.70120","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147972961","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"TOC - Issue Information","authors":"","doi":"10.1111/jmi.70202","DOIUrl":"https://doi.org/10.1111/jmi.70202","url":null,"abstract":"","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":"304 3","pages":""},"PeriodicalIF":1.9,"publicationDate":"2026-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/jmi.70202","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149331308","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Erythrocyte ‘Feierzeit’ reaction: Novel filamentous and vesicular response to n-butyl acetate","authors":"Philip W. Kuchel","doi":"10.1111/jmi.70064","DOIUrl":"10.1111/jmi.70064","url":null,"abstract":"<p>Human erythrocytes (red blood cells; RBCs) undergo spontaneous disassembly after several hours of exposure to <i>n</i>-butyl acetate (<i>n</i>BA). Images of the morphological changes were captured in time-lapse sequences using differential interference contrast (DIC) light microscopy. On exposure to less than 10 mM <i>n</i>BA dramatic events did not take place, but with ∼60 mM aqueous solutions of <i>n</i>BA, discocytes became spherical with a single contiguous ‘geographical’ indentation patch. Over the next ∼2 h the cells became echinocyte-like with rounded projections; and several hours later they discharged filaments that writhed in Brownian motion. In parallel with these changes, vesicles budded from the cells, followed by their alignment on the filaments, like balloons on a string. The vesicles then serially fused, finally giving rise to a single large vesicle that was ∼0.1–0.2 times the diameter of the spherical parent cell; it then fused with the parent cell that a short while later ruptured and became a ‘ghost’. Owing to the striking nature of this phenomenon that was evocative of party decor, the term coined for it was <i>Feierzeit</i> (German: celebration time). The background to this serendipitous discovery, and the deductive odyssey that identified the causative agent, <i>n</i>BA, are presented. Broader implications for understanding the assembly of the RBC cytoskeleton–plasma membrane complexes, and their disassembly under normal, pathological, and forensic conditions are discussed.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":"304 3","pages":"196-208"},"PeriodicalIF":1.9,"publicationDate":"2026-11-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/jmi.70064","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145989337","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}