{"title":"Integrating Electrochemistry and Optical Imaging to Probe Phospholipid Architectures: Emerging Paradigms in Fluorescence and Electrochemiluminescence Studies","authors":"Fatma Ben Trad, Eric Labbé, Olivier Buriez","doi":"10.1021/cbmi.5c00263","DOIUrl":"10.1021/cbmi.5c00263","url":null,"abstract":"<p>The interplay between electrochemistry and optical microscopyisredefining how complex interfacial processes are visualized and quantifiedat the nanoscale. Here, we review and contextualize emerging strategiesthat integrate electrochemical control with fluorescence and electrochemiluminescence(ECL) imaging to probe phospholipid architectures, from supportedlipid bilayers (SLBs) to liposomes or giant unilamellar vesicles (GUVs).In fluorescence-based approaches, the combination of electrochemicalpotential modulation with high-resolution microscopy enables a directcorrelation between redox activity and molecular organization withinmembranes. Recent developments in total internal reflection fluorescencemicroscopy combined with electrochemical fluorescence microscopy (TIRF–ECFM)and redox-sensitive probes have revealed electrochemically inducedlipid reorganization and redox processes within supported bilayerswith unprecedented sensitivity. Complementarily, ECL microscopy extendsthis paradigm by exploiting light emission triggered by electron transferat electrode interfaces, providing label-free and background-freeimaging of lipid film integrity, membrane rupture, and peptide-inducedpermeabilization. Studies on ECL-active liposomes and SLBs demonstratethat variations in emission onset and intensity directly report onthe local membrane composition, permeability, and electrostatics.Moreover, encapsulation of enzymatic + redox-active species withinGUVs enables real-time, noninvasive ECL monitoring of confined biochemicalreactions. Collectively, these emerging electro-optical methodologiesestablish a versatile toolbox for dissecting membrane dynamics andbiochemical reactivity with molecular precision. Their convergenceheralds an original analytical frontier at the interface of electrochemistry,optical imaging, and membrane biophysics, paving the way for next-generationbiosensing, drug screening, and single-vesicle electroanalysis.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 6","pages":"1004–1017"},"PeriodicalIF":6.7,"publicationDate":"2026-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148334332","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}
Zhipeng Shen, Junjie Wen, Xiaohui Ma, Hongxi Zhang, Yi Zhang
{"title":"APT MRI Signature for Risk Stratification of Pediatric Medulloblastoma","authors":"Zhipeng Shen, Junjie Wen, Xiaohui Ma, Hongxi Zhang, Yi Zhang","doi":"10.1021/cbmi.5c00171","DOIUrl":"10.1021/cbmi.5c00171","url":null,"abstract":"<p>Medulloblastoma (MB), a malignant intracranial tumor,is classifiedinto four molecular subgroups (WNT, SHH, Group 3, Group 4) with distinctrisk profiles. This study aimed to establish APT MRI-based radiomicsignature for distinguishing risk subgroups of pediatric MB. FromFebruary 2018 to May 2023, 72 diagnosed pediatric MB patients wereenrolled, all undergoing brain MRI examinations. Radiomic featureswere extracted from regions of interest in APT-related metric mapsand structural MR images, and were subject to a feature selectionprocess. The performance of retained individual features and radiomicsignatures generated via the least absolute shrinkage and selectionoperator (LASSO) was assessed using receiver operating characteristic(ROC) analysis. Final analysis included 56 subjects (mean age: 76.5± 41.9 months; 37 males), consisting of 7 WNT, 17 SHH, 9 Group3, and 23 Group 4 cases. The optimal individual radiomic feature exhibitedan area under the ROC curve (AUC) of 0.83 for risk stratification,while the APT-based radiomic signature achieved a higher AUC of 0.86.Notably, structural images and clinical factors failed to provideadditional value to the APT-based signature for risk stratification.In conclusion, APT MRI-based radiomic signature demonstrated favorableperformance in distinguishing risk groups of pediatric MB, offeringvaluable insights for clinical diagnosis of MB.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 6","pages":"1107–1115"},"PeriodicalIF":6.7,"publicationDate":"2026-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148334336","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}
Davide Panzeri, Reha Akpinar, Laura D’Alfonso, Margaux Bouzin, Luca Presotto, Maddalena Collini, Giuseppe Chirico, Luca Di Tommaso, Donato Inverso, Laura Sironi
{"title":"Multimodal Phasor Analysis for Digital Pathology: Quantitative Characterization of Liver Iron Overload","authors":"Davide Panzeri, Reha Akpinar, Laura D’Alfonso, Margaux Bouzin, Luca Presotto, Maddalena Collini, Giuseppe Chirico, Luca Di Tommaso, Donato Inverso, Laura Sironi","doi":"10.1021/cbmi.5c00229","DOIUrl":"10.1021/cbmi.5c00229","url":null,"abstract":"<p>Hemochromatosis is a pathologicalcondition characterized by theexcessive absorption and storage of iron, primarily in the liver,which can lead to life-threatening complications. Histological tissuesections stained with Perls’ Prussian Blue (PB) are used tovisualize iron accumulation under transmitted light. In clinical practice,pathologists evaluate the density and intensity of these PrussianBlue deposits through direct visual inspection, a semiquantitativeprocess that can be influenced by individual interpretation and experience.Whole-slide scanners now digitize entire tissue sections into high-resolutionRGB whole-slide images (WSIs), allowing image analysis algorithmsto support pathologists with quantitative diagnostics. In this context,we developed a WSI processing method based on a multimodal phasorapproach to quantify the extent, size and distribution of iron deposits,providing quantitative data that closely correlate with pathologists’diagnoses based on the Scheuer scoring system. The Discrete FourierTransform was used to map the RGB spectral data into the phasor plane,where the contribution of the PB stain was isolated using an unsupervisedclustering method. Additionally, the size of Prussian Blue aggregateswas determined through phasor analysis of the spatial autocorrelationfunction. Our results highlight the benefits of phasor-based analysisin segmentation, comprehensive analysis, and the efficient managementof large image data sets. This method has potential applications inanalyzing immunohistochemistry and immunofluorescence spectral images,as well as in simultaneously studying nanoparticle size and diffusionproperties within tissues.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 6","pages":"1186–1195"},"PeriodicalIF":6.7,"publicationDate":"2026-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148334315","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":"FAPI Dimerization for Theranostic Applications: Molecular Design, Preclinical Validation, and Clinical Translation","authors":"Peng Jiang, Kejing Shao, Bao Zhu, Fei Chen","doi":"10.1021/cbmi.6c00005","DOIUrl":"10.1021/cbmi.6c00005","url":null,"abstract":"<p>Fibroblast activation protein (FAP), a serine proteaseoverexpressedin cancer-associated fibroblasts of >90% epithelial malignancies,has emerged as a highly promising pan-cancer target for theranosticradiopharmaceuticals. However, the rapid clearance of monomeric FAPinhibitors (FAPIs) limits their therapeutic efficacy despite excellentdiagnostic performance. Multimerization strategies-particularly dimericand heterodimeric constructs-have been developed to overcome thislimitation through enhanced binding avidity and prolonged tumor retention.This review provides a comprehensive analysis of recent advances inFAPI-based multimers, focusing on their molecular design, chemicalsynthesis, preclinical evaluation, and early clinical applications.We examine the role of linker chemistry (length, flexibility, cleavability)and chelator selection in optimizing pharmacokinetics and tumor-to-backgroundcontrast. Key examples include homodimers (e.g., DOTA-2P(FAPI)<sub>2</sub>, BiOncoFAP) that leverage the polyvalency effect, and heterodimers(e.g., FAPI-RGD, PSFA-01) that enable dual-targeting of FAP and complementaryreceptors such as integrin α<sub>v</sub>β<sub>3</sub> orPSMA. These probes show superior tumor uptake and retention in preclinicalmodels and have demonstrated enhanced diagnostic sensitivity and therapeuticpotential in clinical trials across multiple cancer types. Beyondoncology, emerging applications in fibrotic and inflammatory diseases-suchas rheumatoid arthritis and interstitial lung disease-highlight theversatile utility of FAPI multimers. While challenges including renaluptake, synthetic complexity, and cost remain, ongoing innovationsin chemical design and combination therapies position these agentsas transformative tools in precision theranostics, bridging high-contrastimaging with effective radioligand therapy for personalized patientmanagement.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 6","pages":"1043–1054"},"PeriodicalIF":6.7,"publicationDate":"2026-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148334309","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":"Local Birefringence Imaging by Similar Mueller Matrix Averaging Method in Catheter-Based Polarization-Sensitive Optical Coherence Tomography","authors":"Qingrui Li, Rongyang Zhu, Kun Liu, Junfeng Jiang, Tiegen Liu, Zhenyang Ding","doi":"10.1021/cbmi.5c00203","DOIUrl":"10.1021/cbmi.5c00203","url":null,"abstract":"<p>Local birefringence imaging provides the most intuitivepolarizationproperty of tissue, such as vascular plaque, in catheter-based polarization-sensitiveoptical coherence tomography (PS-OCT) imaging. We present a LocalBirefringence Similar Mueller Matrix Averaging (LMMA) for catheter-basedPS-OCT imaging. LMMA combines differential Mueller analysis, matrixdecomposition, and noise compensation to address depolarization couplingand noise interference. Monte Carlo simulations show LMMA achieves4.8% error under high noise and 4.4% under high depolarization─upto 84% improvement over the Stokes vector spectral binning (SSB) method.Polycarbonate imaging experiments yield 1.5% average error, 86% lowerthan the SSB. <em>Ex-vivo</em> porcine coronary artery imagingdemonstrates significantly enhanced lipid and fibrous plaque phantomdifferentiation, with LMMA’s plaque recognition rate reaching2.62 times that of SSB evaluated by spatial response of Laplacianoperator (SRLO). The results show LMMA’s potential in advancedvascular imaging, particularly for monitoring and diagnosis of coronaryartery disease.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 6","pages":"1163–1172"},"PeriodicalIF":6.7,"publicationDate":"2026-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148334334","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}
Gabriel F. Pelentir, Vanessa R. Bevilaqua, Michio Kakiuchi, Takashi Hirano, Vadim R. Viviani
{"title":"Brighter NIR Bioluminescence System for Mammalian Cell Bioimaging Based on Engineered Railroadworm Luciferase and 6′-Aminoluciferin Analogues","authors":"Gabriel F. Pelentir, Vanessa R. Bevilaqua, Michio Kakiuchi, Takashi Hirano, Vadim R. Viviani","doi":"10.1021/cbmi.5c00163","DOIUrl":"10.1021/cbmi.5c00163","url":null,"abstract":"<p>Luciferaseshave been extensively used as reporter genesfor bioimagingpurposes. For bioimaging in live animals, however, far-red- and NIR-emittingsystems are better suited. Despite many efforts to develop such NIR-emittingsystems, currently only a few systems have been commercialized, includingAkaLuc/akalumine, which emits at ∼650 nm. Departing from thenatural red-emitting luciferase of <em>Phrixotrix hirtus</em> railroadworm in combination with 6′-(1-pyrrolidinyl)luciferin(N5), we previously reported the development of a first efficientfar-red-emitting system (650 nm). Here, we further improved this system,developing a luciferase that emits far-red bioluminescence with thenatural substrate <span>d</span>-luciferin (630 nm), and a more efficientNIR bioluminescence with 6′-amino analogues (650–664nm). The combination of this luciferase with N5 displayed superiorproperties compared with the commercial akalumine/AkaLuc system (645nm), with high affinity for N5, higher catalytic efficiency and thermostability,more sustained luminescence, and more red-shifted spectrum (660 nm).COS-1 fibroblasts expressing this luciferase/N5 combination also displayeda brighter BL signal and more red-shifted spectrum when compared withthe commercial akalumine/AkaLuc system, providing a promising NIRsystem for mammalian BL imaging.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 6","pages":"1098–1106"},"PeriodicalIF":6.7,"publicationDate":"2026-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148334342","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}
Haoyu Niu, Chenhang Wang, Kang Lu, Yixian Wang, Yewei Luo, Xinyan Long, Minrui Xie, Mengze Xu, Sen Yang, Yi Lin, Jiaxin Kuang, Shichao Wang
{"title":"Hypoxia-Activated NIR-II Fluorescent Probes for Breast Tumor Imaging via N-Oxide BF2 Formazanate Scaffolds","authors":"Haoyu Niu, Chenhang Wang, Kang Lu, Yixian Wang, Yewei Luo, Xinyan Long, Minrui Xie, Mengze Xu, Sen Yang, Yi Lin, Jiaxin Kuang, Shichao Wang","doi":"10.1021/cbmi.5c00201","DOIUrl":"10.1021/cbmi.5c00201","url":null,"abstract":"<p>Tumor hypoxia critically influences cancer progressionand therapyresistance, yet its precise and selective visualization remains amajor challenge. In this study, we report a class of small-moleculeNIR-II fluorescent probes (<strong>BFO1</strong>–<strong>BFO3</strong>) based on <em>N</em>-oxide-functionalized boron difluorideformazanates, which are capable of specifically responding to oxygen-deficientenvironments without interference from nitroreductase activity. Theseprobes undergo bioreduction via cytochrome P450-mediated deoxygenationunder hypoxic conditions, leading to a pronounced “turn-on”fluorescence emission beyond 1000 nm. Among them, <strong>BFO1</strong> demonstrated outstanding selectivity, sensitivity, and deep-tissueimaging performance, achieving high-resolution hypoxia detection upto a depth of 5 mm in a 1% intralipid model. Cellular and <em>in vivo</em> studies in 4T1 breast tumor models confirmed therobust and rapid hypoxia-activated NIR-II signal enhancement. Collectively,this work presents an <em>N</em>-oxide-based scaffold forthe noninvasive NIR-II imaging of hypoxia, offering a powerful platformfor early tumor diagnosis and monitoring of hypoxia-associated pathologies.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 6","pages":"1055–1061"},"PeriodicalIF":6.7,"publicationDate":"2026-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148334267","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":"AI-Assisted Coherent Raman Scattering Microscopy for Clinical Translation","authors":"Yue Yu, Yuheng Guo, Kuan Luo, Minbiao Ji","doi":"10.1021/cbmi.5c00199","DOIUrl":"10.1021/cbmi.5c00199","url":null,"abstract":"<p>Coherent Raman scattering (CRS) microscopyhas emerged as a powerfulbioimaging tool, combining bond-specific contrast with high sensitivity,to enable rapid histopathology and metabolic analysis. Yet, clinicaltranslation demands further breakthroughs in acquisition speed, spatial/spectralresolution, imaging depth, and data interpretation. While hardwareadvances address some constraints, artificial intelligence (AI) nowpropels the field forward by decoding biological complexity beyondtraditional limits. This review summarizes representative clinicalapplications of CRS microscopy, highlighting how AI-driven innovationsare bridging technological gaps and accelerating deployment in medicaldiagnostics and research.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 6","pages":"970–983"},"PeriodicalIF":6.7,"publicationDate":"2026-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148334311","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}
Elizabeth J. Fear, Antonella Antonelli, Pasant Abdalla, Isaac Watson, Riccardo Di Corato, Victoria Annis, Oliver J. Mundell, Simon B. Duckett, Luigia Rossi, Elisa Zamboni, Mauro Magnani, Aneurin J. Kennerley
{"title":"Red Blood Cell-Encapsulated Nanoparticles for Long-Circulating, Improved Specificity Functional MRI","authors":"Elizabeth J. Fear, Antonella Antonelli, Pasant Abdalla, Isaac Watson, Riccardo Di Corato, Victoria Annis, Oliver J. Mundell, Simon B. Duckett, Luigia Rossi, Elisa Zamboni, Mauro Magnani, Aneurin J. Kennerley","doi":"10.1021/cbmi.5c00190","DOIUrl":"10.1021/cbmi.5c00190","url":null,"abstract":"<p>Nanoparticle-based MRI contrast agents have shown promisefor advancingnoninvasive imaging, but their clinical utility is limited by rapidclearance, poor biocompatibility, and lack of sustained signal. Here,we present a red blood cell (RBC)-based nanocarrier platform thatencapsulates superparamagnetic iron oxide nanoparticles (SPIONs) followinghypotonic dialysis and resealing of the cell membranes. This biomimetic“Trojan horse” strategy exploits the inherent circulationtime, deformability, and biocompatibility of RBCs to prolong the nanoparticlelifetime and enhance the translational potential. In vivo rodent studiesdemonstrated that SPION-loaded human RBCs provide robust, long-lastingcerebral blood volume (CBV)-weighted functional (f) MRI signal with>5-fold magnitude stronger responses over conventional/establishedblood oxygenation level-dependent (BOLD) contrast. In addition, functionalbrain mapping using cell encapsulated SPIONs show improved laminarspecificity, with activity localized to cortical layer IV. Comparedwith free SPIONs, loaded cells achieved >30 min of stable <em>T</em><sub>2</sub>* contrast at one-quarter of the iron dose,while maintaining physiologically plausible CBV maps. These findingsconfirm efficacy and establish RBC encapsulation as a versatile andbiocompatible nanomedicine platform for extending nanoparticle circulationand enabling high-resolution functional imaging with broad implicationsfor translational applications in neurology, oncology, and theragnostics.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 6","pages":"1139–1152"},"PeriodicalIF":6.7,"publicationDate":"2026-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148334295","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}