Chemical & Biomedical Imaging最新文献

{"title":"An Open, Integrated Platform for Multiplexed Bioluminescence Microscopy","authors":"Lorenzo Scipioni,&nbsp;Belen Torrado,&nbsp;Giulia Tedeschi,&nbsp;Lila P. Halbers,&nbsp;Zachary R. Torrey,&nbsp;Erin B. Fuller,&nbsp;Francesco Fersini,&nbsp;Christoph Gohlke,&nbsp;Andrej Luptak,&nbsp;Jennifer A. Prescher,&nbsp;Michelle A. Digman","doi":"10.1021/cbmi.5c00225","DOIUrl":"https://doi.org/10.1021/cbmi.5c00225","url":null,"abstract":"<p>We report an imaging package to democratize all-in-onebioluminescenceand fluorescence microscopy. The platform comprises three tools: PhasorViewer,a visualization suite to design experiments and identify optimal probecombinations; PhasorScope, an open-source, cost-effective microscopyframework to upgrade conventional microscopes; and PhasorAnalysis,a dedicated, user-friendly analysis pipeline to quantify phasor imagingdata. We demonstrate the utility of the platform for multiplexed,simultaneous fluorescence and bioluminescence imaging with readilyaccessible optical reporters.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 9","pages":"2003–2009"},"PeriodicalIF":6.7,"publicationDate":"2026-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149067469","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multimodal Biophysical Analysis of Morpho-Phenotypic and Morpho-Functional Dynamics in Normal and Cancer Spheroids within Engineered Microenvironments","authors":"Srijani Dey,&nbsp;Shreya Adhikary,&nbsp;Priyanka Das,&nbsp;Pallab Datta,&nbsp;Subhadip Chakraborty,&nbsp;Ananya Barui","doi":"10.1021/cbmi.5c00255","DOIUrl":"https://doi.org/10.1021/cbmi.5c00255","url":null,"abstract":"<p>Three-dimensional spheroid models provide physiologicallyrelevantsystems to investigate cancer progression and stem cell dynamics.However, most existing literature focuses primarily on morphologicalor biochemical characterization, limiting a comprehensive understandingof the coupled biophysical, morphological, and molecular dynamicsof spheroids within engineered microenvironments. Addressing thisgap, the present study develops a multimodal biophysical frameworkto comprehensively assess the morpho-phenotypic, metabolic, and electrochemicalcharacteristics of spheroids. Here, we report a comparative analysisof spheroids derived from mesenchymal stem cells (MSC), a mixed populationof cancer stem cells and cancer cells (A549), and differentiated cancercells (MCF-7), cultured in an ECM-mimetic alginate-gelatin hydrogel.Relevant morpho-phenotypic parameters were analyzed at 3, 7, and 14days from confocal images and cytoskeletal mapping, revealing distincttrends in size, circularity, solidity, and entropy. All spheroidsexhibited time-dependent growth (40–60 to 120–240 μm),with cancer spheroids (A549 and MCF-7) showing significantly greatervolumetric expansion. Day 7 emerges as a critical transition pointmarked by an alteration in the morphological trend. A549 spheroidsdisplayed higher entropy, reflecting increased heterogeneity. Autofluorescencespectroscopy revealed distinct redox dynamics of spheroids from differentorigins. Cancer spheroids showed enhanced NADH, FAD, and porphyrinsignatures reflecting redox alterations and Warburg-like effects,wherein MSC spheroids showed a controlled shift toward oxidative phosphorylation.The impedance spectroscopy further distinguished spheroids throughcapacitance–resistivity relationships. Notably, A549 spheroidsexhibit dynamic transitions between stem-like and malignant states,consistent with their intrinsic heterogeneity. Overall, this integratedapproach correlates growth, redox status, and dielectric behavior,offering a translational platform for drug screening, disease modeling,and regenerative medicine.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 9","pages":"2010–2022"},"PeriodicalIF":6.7,"publicationDate":"2026-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149067474","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"100-Second Large-Area Chemical Imaging by Integrated Four-Channel CARS Microscopy","authors":"Wei Ju,&nbsp;Zhiliang Huang,&nbsp;Shangyu Li,&nbsp;Yunshan Zhong,&nbsp;Yiran Li,&nbsp;Na Li,&nbsp;Chunxian Yang,&nbsp;Yue Huang,&nbsp;Shuai Wang,&nbsp;Shuai Yan,&nbsp;Jiangling Wan,&nbsp;Xiangliang Yang,&nbsp;Ping Wang","doi":"10.1021/cbmi.6c00016","DOIUrl":"https://doi.org/10.1021/cbmi.6c00016","url":null,"abstract":"<p>Label-free coherent Raman scattering(CRS) microscopyenables chemicalobservation of intricate cellular and intercellular structures ofbiological tissues with submicron resolution and has become the mostprominent stain-free alternative to gold-standard hematoxylin andeosin (H&amp;E) histology. However, the imaging field of view (FOV)of current CRS microscopes is typically limited to less than a millimeter.Various attempts in instrumentation have been made to perform large-scaleCRS mapping across an intact tissue, but the costly trade-offs areeither imaging speed or spatial resolution. To address this challenge,we developed a four-channel coherent anti-Stokes Raman scattering(CARS) microscope, offering submicron imaging of 13 × 13 mm² large tissues within 100 s. Four miniaturized CARS systems,each equipped with a microelectromechanical system (MEMS) scanner,a micro-objective, and a collection fiber bundle, were fully integratedto perform ultrafast chemical mapping of intact unprocessed tissueswith an electronically synchronized motorized stage. We demonstratedits capability by performing large-scale CARS imaging of mouse braintissues and human glioma tissues. This label-free technique may pavethe way for future clinical applications of stain-free pathology inthe operating room.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 9","pages":"2084–2090"},"PeriodicalIF":6.7,"publicationDate":"2026-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149067489","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Quantitative Analysis of Cytoplasmic Viscosity in Colorectal Cancer Cells by Differential Dynamic Microscopy of Genetically Encoded Nanoparticles","authors":"Steven Huysecom,&nbsp;Kiki Schouwstra,&nbsp;Samet Aytekin,&nbsp;Natalie Neyrinck,&nbsp;Yana Heyvaert,&nbsp;Boris Louis,&nbsp;Susana Rocha,&nbsp;Guillermo Solís-Fernandez","doi":"10.1021/cbmi.5c00261","DOIUrl":"https://doi.org/10.1021/cbmi.5c00261","url":null,"abstract":"<p>The viscosity of the cytoplasm plays a key role in regulatingmoleculardiffusion and cellular mechanics, yet quantifying it in living cellsremains technically challenging. Genetically encoded multimeric nanoparticles(GEMs) have emerged as useful probes for intracellular microrheology;however, current analyses rely on single-particle tracking, whichis limited by probe density, imaging noise, and expression variability.Here, we combine GEMs with differential dynamic microscopy (DDM) toenable quantitative, noninvasive, and rapid measurement of intracellularviscosity using standard wide-field fluorescence imaging. DDM extractsparticle dynamics from ensemble spatiotemporal intensity fluctuations,yielding diffusion coefficients and viscosity estimates, even in crowdedor heterogeneous environments where tracking fails. Validation withfluorescent nanoparticles diffusing in water confirmed that DDM accuratelyreproduced theoretical viscosities across a wide range of particlesizes and concentrations. Comparison with single-particle tracking(SPT) demonstrated equivalent precision under dilute conditions andimproved the performance under crowding. To showcase the potentialof this approach, we applied GEM-DDM to colorectal cancer cell lineswith different metastatic potentials. Cytoplasmic viscosity correlatedwith aggressiveness, increasing from 1.9 to 2.3 cP in poorly metastaticlines to 3.6–3.7 cP in highly metastatic lines, consistentwith greater macromolecular crowding and cytoplasmic reorganizationreported in aggressive cells. Together, these results establish GEM-DDMas a fast, reproducible, and accessible platform for intracellularmicrorheology to link the physical state of the cytoplasm to cellfunction and disease progression.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 9","pages":"1978–1986"},"PeriodicalIF":6.7,"publicationDate":"2026-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149067496","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effects of Ligand Mobility on Ligand–Receptor Interactions Driving Integrin-Mediated Cell Adhesion","authors":"Eunseo Cho,&nbsp;Jaeyi Hahm,&nbsp;Jaekyung Moon,&nbsp;Young-Jin Seo,&nbsp;Hye Ran Koh","doi":"10.1021/cbmi.6c00057","DOIUrl":"https://doi.org/10.1021/cbmi.6c00057","url":null,"abstract":"<p>Ligand–receptor interactions that drive cell adhesionoccurwithin dynamic membrane environments, where ligand mobility can influenceforce transmission and downstream signaling. Despite its potentialsignificance, the mechanical role of ligand mobility in regulatingthe activation threshold for cell adhesion remains poorly understood.Here, we quantified how ligand mobility modulates the apparent adhesionthreshold for integrin-mediated cell adhesion using DNA-based tensiongauge tethers. By directly comparing mobile ligands presented on supportedlipid bilayers with immobilized ligands on rigid substrates, we demonstratedthat mobile ligands exhibited apparent adhesion thresholds consistentlylower than those of immobile ligands across multiple cell types. Controlledmodulation of membrane fluidity further revealed that the ligand diffusionconstants are inversely proportional to apparent adhesion thresholds.These results highlight the critical role of ligand mobility in ligand–receptorinteractions and advance our understanding of how membrane physicalproperties regulate cellular signaling.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 9","pages":"1996–2002"},"PeriodicalIF":6.7,"publicationDate":"2026-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149071078","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cell Death and Unprecedent Cytoskeletal Changes in Leishmania amazonensis Parasites Evidenced by 3D Reconstruction","authors":"Rayanne Regina Beltrame Machado,&nbsp;Amanda Beatriz Kawano Bakoshi,&nbsp;Tânia Ueda-Nakamura,&nbsp;Sueli de Oliveira Silva Lautenschlager,&nbsp;Maria Helena Sarragiotto,&nbsp;Danielle Lazarin-Bidóia,&nbsp;Celso Vataru Nakamura","doi":"10.1021/cbmi.6c00109","DOIUrl":"https://doi.org/10.1021/cbmi.6c00109","url":null,"abstract":"<p>Leishmaniasis remains a neglected tropical disease withlimitedand highly toxic therapeutic options. In search of safer and moreeffective alternatives, bithiophene derivatives have emerged as promisingscaffolds. In this study, we evaluated the antileishmanial potentialof <strong>BT-Ac</strong>, a bithiophene previously shown to exert antitrypanosomalactivity. <strong>BT-Ac</strong> displayed selective antiproliferativeactivity with IC₅₀ for 28.9 and 50 μM against promastigotesand amastigotes of <em>Leishmania amazonensis</em> at nontoxic concentrations for host cells, achieving a selectivityindex of 4.69 against intracellular amastigotes. Electron microscopyrevealed early morphological damage in promastigotes, including unprecedentedchanges at the plasma membrane and cytoskeletal interface, mitochondrialswelling, lipid body accumulation, and autophagic vacuole formation.Three-dimensional reconstructions further validated these findings,confirming that the ultrastructural alterations were intrinsic to <strong>BT-Ac</strong> treatment rather than technical artifacts. Biochemicalanalyses demonstrated reactive oxygen and nitrogen species overproduction,lipid peroxidation, and ATP fluctuations, indicating oxidative stressas a central mechanism. These events culminated in apoptosis-likeand autophagic death in promastigotes, while amastigote death wasprimarily mediated by macrophage activation, evidenced by nitric oxiderelease. Together, our findings identify <strong>BT-Ac</strong> as a selectiveleishmanicidal agent that disrupts parasite homeostasis through oxidativedamage, highlighting its potential as a lead compound for therapeuticstrategies against leishmaniasis.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 9","pages":"2102–2118"},"PeriodicalIF":6.7,"publicationDate":"2026-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149067454","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pathogen-Specific PET Imaging of Pulmonary Infection Using [18F]Fluoromannitol","authors":"Spenser R. Simpson,&nbsp;Tyler Simmons,&nbsp;Haley Echlin,&nbsp;Shyra Wilde,&nbsp;Shashika S. Perera,&nbsp;Paulina Villanueva,&nbsp;Amy Iverson,&nbsp;Amber Schuster,&nbsp;Peter Vogel,&nbsp;Amanda Green,&nbsp;Jason Rosch,&nbsp;Kiel D. Neumann","doi":"10.1021/cbmi.5c00303","DOIUrl":"https://doi.org/10.1021/cbmi.5c00303","url":null,"abstract":"<p>Pneumonia remains a leading cause of global morbidityand mortality,with diagnostic uncertainty frequently complicating clinical management,particularly in patients with chronic lung disease, immunocompromise,or healthcare-associated infections. Conventional imaging modalitiesand microbiologic testing lack specificity for distinguishing bacterialpneumonia from viral infection, sterile inflammation, or noninfectiouslung disease, often resulting in empiric antibiotic overuse and delayedalternative therapies. In this study, we evaluated [¹⁸F]fluoromannitol,a pathogen-specific Positron Emission Tomography (PET) tracer, inwell-established murine models of pulmonary infection caused by <em>Streptococcus pneumoniae</em>, <em>Staphylococcus aureus</em>, and <em>Pseudomonas aeruginosa</em>. Compared with controls, including heat-killed bacterial pneumonitis,influenza A virus–induced pneumonia, and noninfectious Poly(I:C)induced pneumonia. [¹⁸F]fluoromannitol PET enabled robust,rapid, and anatomically localized detection of active bacterial pneumonia,with tracer uptake correlating with bacterial burden and disease progression.These findings demonstrate that [¹⁸F]fluoromannitol PETcan noninvasively distinguish bacterial lung infection from viraland noninfectious etiologies. By directly targeting bacterial metabolismrather than host inflammation, this approach offers a promising strategyto improve etiologic classification of lower respiratory tract infectionsand support antimicrobial stewardship.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 9","pages":"2074–2083"},"PeriodicalIF":6.7,"publicationDate":"2026-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149067483","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mechanisms and Applications of Photocatalytic Proximity Labeling","authors":"Enze Gong,&nbsp;Yalan Xie,&nbsp;Wei Qin","doi":"10.1021/cbmi.5c00276","DOIUrl":"https://doi.org/10.1021/cbmi.5c00276","url":null,"abstract":"<p>Photocatalytic proximity labeling(PPL) is a light-controlled techniquefor spatially and temporally precise mapping of biomolecular interactions.This review classifies PPL platforms into small-molecule catalysts(transition metal complexes and organic chromophores) and geneticallyencoded photosensitizers, detailing their mechanisms and biologicalapplications─from subcellular proteomics, cell–cellinterface tagging to in vivo labeling. We compare PPL with enzyme-catalyzedmethods, highlighting its advantages in spatiotemporal control, minimalgenetic manipulation, and suitability for primary cells. Current challengesin PPL are also discussed. As PPL methodologies continue to evolvefrom mechanistic innovation toward essential biological problem-solving,they hold great promise for elucidating dynamic interaction networkswith unprecedented resolution in living systems.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 9","pages":"1915–1932"},"PeriodicalIF":6.7,"publicationDate":"2026-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149067495","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mapping Nanoscale Forces and Potentials in Live Cells with Microsecond 3D Single-Particle Tracking","authors":"Shangguo Hou,&nbsp;Chen Zhang,&nbsp;Anastasia Niver,&nbsp;Kevin D. Welsher","doi":"10.1021/cbmi.5c00293","DOIUrl":"https://doi.org/10.1021/cbmi.5c00293","url":null,"abstract":"<p>3D single-particle tracking has the potential to resolvethe molecular-levelforces that dictate particle motion in biological systems. However,the information gleaned from 3D single-particle tracking often cannotresolve underlying nanoscale potentials due to limited spatiotemporalresolution. To this end, we introduce an active-feedback 3D trackingmicroscope that utilizes silver nanoparticles (AgNPs) as probes tostudy intricate biophysical events in live cells at the nanometerand microsecond scales. Due to the extremely high, durable scatteringphoton flux of the plasmonic particles, a 1 MHz sampling frequencywith nanometer precision in all three dimensions can be achieved overan unlimited observation duration. In this work, microsecond-sampling,active-feedback 3D single-particle tracking was applied to investigatethe interaction between AgNPs and nanoscale filopodia on the live-cellsurface. The nanometer precision and microsecond sampling revealedthat TAT peptide-modified particles visit and dwell at local “hotspots” on the filopodium surface. The high sampling rate furtherenabled the calculation of local forces and potentials within thesenanoscale hotspots on the cylindrical surfaces of live-cell filopodia.This study presents a promising tool for investigating intracellularbiophysical events with unprecedented spatiotemporal resolution, alongwith a pipeline for studying nanoscale potentials within three-dimensionalcellular structures.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 9","pages":"2054–2064"},"PeriodicalIF":6.7,"publicationDate":"2026-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149067471","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Donor–Acceptor Bridging via Twisted Quasi-Double-Bond: A Facile Design Strategy for Typical NIR-II AIEgen with “Simple” Structure","authors":"Chunbin Li,&nbsp;Jen-Shyang Ni,&nbsp;Pengfei Zhang,&nbsp;Daoyong Jiang,&nbsp;Jianguo Wang,&nbsp;Kai Li,&nbsp;Shuang-Quan Zang,&nbsp;Ryan T. K. Kwok,&nbsp;Weihong Tan,&nbsp;Ben Zhong Tang","doi":"10.1021/cbmi.5c00302","DOIUrl":"https://doi.org/10.1021/cbmi.5c00302","url":null,"abstract":"<p>A facile design strategy for NIR-II fluorophores withaggregation-inducedemission (AIE) characteristics was presented. This kind of NIR-IIAIE luminogen, TSPCI is an example, is by the strong acceptor–donorbridging via a twisted quasi-double-bond (TQDB). Theoretical calculationsrevealed that the introduction of TQDB was the major factor for quenchingof the photoexcited TSPCI molecule in the solution state. TSPCI showedexcellent NIR-II imaging performance for in vivo dynamic imaging withhigh contrast and penetration depth. TSPCI holds great potential asa building block to easily fabricate activatable fluorescent probeswith superior responsive ability in the second NIR optical windowfor in vivo imaging applications.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 9","pages":"1987–1995"},"PeriodicalIF":6.7,"publicationDate":"2026-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149067494","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}