Chemical & Biomedical Imaging最新文献

{"title":"Beveled Fiber Probe for Improving Spectroscopic Signal Sensing from Bowel Mucosa Tissue","authors":"Omer Karaborklu,&nbsp;Venkataraman Subramanian,&nbsp;Animesh Jha","doi":"10.1021/cbmi.5c00120","DOIUrl":"10.1021/cbmi.5c00120","url":null,"abstract":"<p>The goal of this work is to devisean optical fiber probe configurationfor fiber fluorescence spectroscopy to improve its diagnostic capabilityin gastrointestinal (GI) cancers. Proposed design upgrades to theprobe’s distal tip showed a major increase in spectroscopicsignal sensing compared to standard probes, indicating that the accuracyof the method may be improved significantly. GI cancers are globallycommon and mostly seen in the colon, stomach, and esophagus, wherecurrent diagnosis is made using endoscopy and histopathology. However,endoscopy cannot visualize early signs of cancer, while histopathologyrequires a biopsy and takes several weeks to conclude. Fiber spectroscopymay provide rapid and robust diagnostics at early stages in vivo orex vivo and may be able to guide clinical decisions to further curtailthe risk of advancing to later stages of cancer. However, its medicaldiagnostic performance has not reached the gold standard level yetand, thus, needs to be studied further to improve its accuracy. Therefore,we demonstrate the application of a beveled probe tip and specificfiber positioning in a combinatorial approach and experimentally comparethe design with several standard probes to assess their efficiencies.Results show that the proposed probe is notably more efficient incollecting fluorescent signals from excised small human colon tissues.Findings of the current study may help in implementing an alternativerapid path for histopathological standardization ex vivo for unambiguousdecision making and may enhance the precision of endoscope-adaptedin vivo fiber spectroscopy to provide real-time and noninvasive earlydiagnosis in GI cancers.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 7","pages":"1370–1378"},"PeriodicalIF":6.7,"publicationDate":"2026-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148622537","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":"Application of Cumulative Threshold Approaches to Tissue Iron Measurement on Histochemical or Synchrotron X-ray Fluorescence Platforms","authors":"Chan-An Lin,&nbsp;Elvis Acquah,&nbsp;Jake Brooks,&nbsp;Joanna F. Collingwood,&nbsp;Daniel M. Johnstone,&nbsp;Adrienne E. Milward,&nbsp;Rebecca J. Hood","doi":"10.1021/cbmi.5c00230","DOIUrl":"10.1021/cbmi.5c00230","url":null,"abstract":"<p>Digitized imaging of the spatial distribution of a targetedmetalover a given area converts a continuous spectrum of data into a setof discrete pixels of intensities that ideally correlate stronglywith the areal density map. This should be independent of the metalimaging modality, allowing semiquantitative analysis. In practice,correlation strength may vary between data sets obtained by differentmodalities. One enduring problem for all modalities is selecting acutoff threshold intensity appropriately distinguishing the true signalfrom background noise. This may entail subjective choices between“conservative” thresholds prioritizing specificity oversensitivity (i.e., true positives despite possible false negatives)and “discovery-driven” thresholds prioritizing identificationof putative effects for subsequent validation (i.e., true negativesdespite possible false positives). Computerized data processing mayhelp make this more objective by comparing outcomes for the set ofall possible threshold values, termed cumulative threshold analysis.We address pitfalls in performing valid cumulative threshold analysisusing ImageJ/Fiji for relative quantification of brain iron in micewith normal or genetically elevated iron, assessed by classical histochemistryor synchrotron X-ray fluorescence. In addition to pitfalls in choosingsettings for generating analysis histograms, these include data losswith conversion between bit depths and selecting both appropriate <em>X</em>-axis directionality and image display range minima andmaxima for analyzing multiple images. If these factors are handledappropriately, cumulative thresholding provides a powerful approachfor more objective analysis of biometal imaging. This has importantapplications for metal imaging in research and clinical settings.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 7","pages":"1416–1431"},"PeriodicalIF":6.7,"publicationDate":"2026-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148622560","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":"A Hierarchical Etched Grid for Correlative Chemical Imaging","authors":"Natalie S. W. Arigundiya,&nbsp;Jake Brooks,&nbsp;Peter Sykes,&nbsp;Seojin Lee,&nbsp;Naomi P. Visanji,&nbsp;Christopher M. Morris,&nbsp;Kalotina Geraki,&nbsp;Joanna F. Collingwood","doi":"10.1021/cbmi.5c00231","DOIUrl":"10.1021/cbmi.5c00231","url":null,"abstract":"<p>Correlative microscopy linking synchrotron X-ray fluorescence(SXRF)with optical imaging is valuable for contextualizing chemical elementdistributions in biology. The spatial correlation necessary to achievethis presents fundamental challenges and can be a significant constrainton accuracy and data interpretation. We present a technical solutionbased on a finder grid concept, optimized for SXRF correlative studiesof metals in biological tissues, with scope for wider adaptation andapplication. A hierarchically patterned fiducial system was directlyetched onto spectroscopically clean quartz substrates via femtosecondlaser ablation. This design enables improved correlation among SXRF,optical imaging, and histological staining over a greater range oflength scales than conventional registration methods such as the useof tissue architecture from serial sections and the use of electron-microscopy-resolutionfinder grids and applied fiduciary markers that can introduce XRF-signal-dominatinglevels of elements such as copper, nickel, gold, and titanium. Wepresent two quartz finder grid formats: a microgrid and a nanogriddesign. We demonstrate their utility for rapid ROI relocalizationand same-section correlative workflows using human brain tissue. Theetched quartz finder grid approach facilitates rapid and reproducibleROI relocalization and alignment across instruments, particularlywhere integral fiducial markers are sparse or ambiguous.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 7","pages":"1538–1546"},"PeriodicalIF":6.7,"publicationDate":"2026-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148622549","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":"Unraveling the Hidden Balance between Quantum Confinement and Structural Modifications in Near-Infrared Fluorescent Carbon Dots","authors":"Muhammad Madni,&nbsp;Zia Ullah,&nbsp;Hanfang Jiang,&nbsp;Jhilik Roy,&nbsp;Yibin Yan,&nbsp;Dongxiang Chen,&nbsp;Zhengyang Jin,&nbsp;Xihao Wu,&nbsp;Yuante Lan,&nbsp;Shaohua Wang,&nbsp;Shubham Roy,&nbsp;Bing Guo","doi":"10.1021/cbmi.5c00234","DOIUrl":"10.1021/cbmi.5c00234","url":null,"abstract":"<p>Near-infrared (NIR) fluorescent carbon dots (CDs) haveemergedas promising nanomaterials for advanced bioimaging, sensing, and theranosticapplications due to their intrinsic biocompatibility, photostability,tunable emission, and structural versatility. However, achieving strong,stable, and predictable NIR emission remains a major challenge dueto the complex interplay between quantum confinement, surface states,and structural modifications. From a synthesis standpoint, leveragingwell-defined molecular precursors, precisely controlled heteroatomdoping, and defect engineering provides a systematic approach to modulateemissive centers and achieve reproducible, high-performance NIR fluorescence.This review unravels the hidden balance between these governing factorsand provides a unified mechanistic perspective on how sp²/sp³ domain engineering, heteroatom doping, defect modulation,and donor–acceptor interactions collectively dictate long-wavelengthemission. The synthetic routes ranging from top-down to bottom-upapproaches have been discussed comprehensively to highlight how precursorchemistry, reaction environment, and postsynthetic processing influencethe electronic structure and photophysical behavior of CDs. Despitesignificant progress in optimizing the functionality of CDs, key barriers,including inconsistent synthesis, unresolved emission mechanisms,and limited translational readiness, continue to hinder clinical deployment.By bridging fundamental photo physics with emerging biomedical andoptoelectronic applications, this review outlines critical designprinciples and future directions for the rational development of next-generationNIR-emissive CDs.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 7","pages":"1214–1247"},"PeriodicalIF":6.7,"publicationDate":"2026-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148622593","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":"Computed Tomography Imaging and Characteristics of In Situ Forming Implants with Different PLGA Endcaps","authors":"Xinhao Lin,&nbsp;Zixuan Zhen,&nbsp;Seyyed Majid Eslami,&nbsp;Nour Al Zouabi,&nbsp;Lauren Elizabeth Ward,&nbsp;Mittal Darji,&nbsp;Sheyda Ranjbar,&nbsp;Francis K. Masese,&nbsp;André O’Reilly Beringhs,&nbsp;Rajeswari M. Kasi,&nbsp;Qiangnan Zhang,&nbsp;Qi Li,&nbsp;Qin Bin,&nbsp;Yan Wang,&nbsp;Hong Yuan,&nbsp;Xiuling Lu","doi":"10.1021/cbmi.6c00001","DOIUrl":"10.1021/cbmi.6c00001","url":null,"abstract":"<p>A poly(lactic-<em>co</em>-glycolic acid) (PLGA)-basedinsitu-forming implant (ISFI) is a long-acting injectable composed ofactive ingredients, a biodegradable polymer and a biocompatible solvent.Understanding the role of PLGA end-caps is crucial for designing ISFIformulations due to their impact on drug delivery performance. Inthis study, using leuprolide acetate as a model drug, we characterizedthe ISFIs using computed tomography (CT) imaging to explore the influenceof PLGA end-cap on the implant’s formation and drug-releasebehavior. CT imaging enabled a detailed characterization of implantmorphology and internal structure, providing new insights into therelationship between phase inversion, implant formation, and in vitrorelease performance. Acid-ended PLGA showed a higher initial burstrelease with 100% of the solvent released at 9 days and a faster releaseduration of the drug in the in vitro release profile. In contrast,ester-ended PLGA showed a more prolonged release profile, with theplateau phase not reached until approximately day 50. Morphologicalanalysis from CT images of the in vitro implants revealed that acid-endedPLGA formed spherical implants with a dense outer layer, whereas ester-endedPLGA resulted in irregular shapes with a larger size expansion. Invivo CT imaging confirmed these trends, although implant evolutionoccurred more rapidly in biological environments. Overall, this researchhighlights the impact of PLGA end-caps on the performance of ISFI,providing a scientific basis for formulation development, evaluation,and optimization of ISFIs.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 7","pages":"1509–1520"},"PeriodicalIF":6.7,"publicationDate":"2026-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148622517","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":"Specific PET Imaging for Precision Management of Lung Cancer: Advances, Clinical Translation and Future Directions","authors":"Chongyang Chen,&nbsp;Donghui Pan,&nbsp;Xinyu Wang,&nbsp;Yuping Xu,&nbsp;Junjie Yan,&nbsp;Lizhen Wang,&nbsp;Min Yang","doi":"10.1021/cbmi.5c00289","DOIUrl":"10.1021/cbmi.5c00289","url":null,"abstract":"<p>Lung cancer continues to be the leading cause of cancer-relateddeaths worldwide, primarily due to persistent challenges in earlydetection and the limited effectiveness of precision medicine. Althoughlow-dose computed tomography (CT) has been widely implemented forlung cancer screening and has contributed to a measurable reductionin disease-specific mortality, its diagnostic accuracy is limitedby its inability to reliably distinguish benign from malignant pulmonarynodules. Furthermore, the clinical standard for metabolic imaging─¹⁸F-fluorodeoxyglucose (¹⁸F-FDG) positron emissiontomography (PET)─is significantly constrained by a high incidenceof false-positive findings in patients with inflammatory conditionsand frequent false-negative results in tumors exhibiting low glycolyticactivity. To overcome these critical limitations, highly specificPET radiotracers have been developed to target immune checkpoints,cell surface receptors, and distinct features of the tumor microenvironment.These molecularly targeted probes offer improved biological specificity,enabling more precise visualization of tumor pathophysiology and supportingapplications in early diagnosis, real-time treatment monitoring, andprognostic stratification. This review summarizes recent advancesin targeted PET imaging and critically evaluates the potential ofintegrating these approaches with multimodal imaging, liquid biopsy,and artificial intelligence (AI)-driven radiomics to enhance diagnosticaccuracy and inform therapeutic decision-making. By synthesizing keydevelopments from both preclinical studies and clinical trials, wehighlight the translational significance and future directions oftarget-specific PET tracers. Despite existing challenges related totracer development, regulatory approval, and methodological standardization,the integration of next-generation whole-body PET systems with advancedartificial intelligence algorithms holds considerable promise forestablishing molecular-targeted PET as a cornerstone of precisiononcology in lung cancer management.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 7","pages":"1293–1314"},"PeriodicalIF":6.7,"publicationDate":"2026-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148622581","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":"Fast, Streamlined In Situ Genotyping of Bacterial Strains for Single-Cell and Subcellular Dynamics Profiling","authors":"Wenjie Zhang,&nbsp;Yanqing Cai,&nbsp;Mason Hong Cheung,&nbsp;Ying Xu,&nbsp;Angela Ruohao Wu,&nbsp;Yi Liao","doi":"10.1021/cbmi.5c00220","DOIUrl":"10.1021/cbmi.5c00220","url":null,"abstract":"<p>Temporal dynamics, such as cell-cycle oscillations, transcriptionalbursting, and the regulated assembly and disassembly of phase-separatedcondensates, constitute an important layer of regulation in biology.Yet, existing technologies, including DNA sequencing, plate readers,and fluorescence microscopy, face a fundamental trade-off betweentemporal resolution, single-cell and subcellular information, andexperimental throughput. This trade-off not only fragments our understandingof fundamental cellular processes but also impedes large-scale screeningfor dynamic phenotypes. Efforts to mitigate such limitations are beginningto emerge. One promising approach combines live-cell time-lapse imagingof pooled bacterial libraries in microfluidic chips with sequentialfluorescence <em>in situ</em> hybridization (seqFISH) to connectdynamic phenotypes exhibited by individual cells with their correspondinggenotypes. However, widespread adoption of this strategy has beenconstrained by two technical challenges: (1) compromised <em>in situ</em> genotyping efficiency due to nonspecific fluorescentprobe aggregation in confined microfluidic growth chambers, and (2)the reliance on high-copy-number plasmids to ensure genotyping sensitivity,which often distorts dynamic phenotypes and perturbs normal physiologyduring live-cell imaging. Here, we address both limitations with anintegrated and physiologically compatible solution. We introduce asimple surface-modification strategy to suppress fluorescent probeaggregation in microfluidic chips, enabling robust <em>in situ</em> genotyping to be completed overnight. In parallel, we implementbarcoded copy-number-tunable plasmids to decouple the conflictingrequirements of live-cell imaging and post hoc genotyping, therebypreserving delicate subcellular dynamics while maintaining genotypingsensitivity. Together, these improvements transform image-based bacterialscreening into a faster and minimally perturbative workflow, expandingthe scalability and versatility of high-throughput single-cell andsubcellular dynamics screening for both fundamental and syntheticbiology research.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 7","pages":"1405–1415"},"PeriodicalIF":6.7,"publicationDate":"2026-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148622465","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":"Nanoscale-Sensitive VSFG Microscopy for Identifying Breast Tumor in FFPE Samples","authors":"Jianyu Ren,&nbsp;Chun-Chieh Yu,&nbsp;Aida Mestre-Farrera,&nbsp;Elizabeth A. Pfau,&nbsp;Jing Yang,&nbsp;Wei Xiong","doi":"10.1021/cbmi.5c00240","DOIUrl":"10.1021/cbmi.5c00240","url":null,"abstract":"<p>Nanoscale collagen remodeling isa critical physicalsignatureof tumor development and metastasis. Recently, we have shown thatvibrational sum-frequency generation (VSFG) microscopy can detecttumor tissues prepared from Optimal Cutting Temperature (OCT) samples,where this unique sensitivity to nanostructure comes from mode-specificcoherent interference. Yet, most clinical tissues are processed asformalin-fixed, paraffin-embedded (FFPE) blocks, which remains unknownwhether these harsh fixations and embedding processes destroy thesubtle structural cues that VSFG detects. Thus, it could present alarge barrier for further application of VSFG broadly into biophysicsand biomedical research of tumors. Here, we demonstrate that VSFGremains fully effective in deparaffinized FFPE tissues. Crucially,following deparaffinization, key diagnostic metrics were statisticallyindistinguishable from OCT cryosection controls. These metrics includedVSFG spectra, chemical images, and the collagen ratio <em>I</em>_<em>NHs</em>/<em>I</em>_{<em>CH₂</em>}_,<em>Ss</em>. Complementaryatomic force microscopy (AFM) nanomechanics corroborated this finding.These findings open the path of using VSFG imaging for standard clinicalworkflows and the vast global archives of FFPE tissues for retrospectiveprognostic studies and label-free diagnostics.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 7","pages":"1352–1360"},"PeriodicalIF":6.7,"publicationDate":"2026-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148622463","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":"Lipid-Targeting Multiresponsive Benzothiadiazole Fluorophore for Live-Cell Imaging and Real-Time Liver Injury Monitoring","authors":"Brenno A. D. Neto,&nbsp;Fabiano S. Rodembusch,&nbsp;Jose Raimundo Correa,&nbsp;Raissa Kelly Correa de Paiva,&nbsp;Jenny Esmeralda Padilla Sorto,&nbsp;Luciana Machado Ramos,&nbsp;Lilian C. da Luz,&nbsp;Luana Lemos Reis,&nbsp;Paulo Eduardo Narcizo de Souza,&nbsp;Daniel Francisco Scalabrini,&nbsp;Thaís Moura,&nbsp;Claudia C. Gatto,&nbsp;Wanderson Ferreira,&nbsp;Gustavo B. Menezes","doi":"10.1021/cbmi.5c00242","DOIUrl":"10.1021/cbmi.5c00242","url":null,"abstract":"<p>This work describes the multiresponsive photophysicalbehaviorof <strong>BTD-4AS</strong>, a donor–acceptor benzothiadiazolefluorophore. <strong>BTD-4AS</strong> shows pronounced solvatochromism,viscosity- and temperature-dependent emission, and large Stokes shifts,consistent with a twisted intramolecular charge transfer state supportedby Lippert-Mataga and Reichardt analyses. Its fluorescence is stronglypH-sensitive (maximum near pH ≈6–8) and markedly enhancedin micellar and viscous media. The dye’s photostability andlipophilicity support its use as a versatile probe for polarity, viscosity,pH, and lipid-rich environments. Building on these properties, <strong>BTD-4AS</strong> enables selective cellular bioimaging of lipid dropletsand, in a mouse model, reports liver injury through polarity/viscosity-dependentspectral shifts─red emission in damaged regions exposed toextracellular milieu and green emission in intact lipid-rich cells.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 7","pages":"1432–1454"},"PeriodicalIF":6.7,"publicationDate":"2026-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148622558","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":"Linking Adjuvant Immune Stimulation to Spatiotemporal Dynamics via Cross-Scale Photoacoustic Microscopy","authors":"Dong Liu,&nbsp;Fengbing He,&nbsp;Yu Zhang,&nbsp;Yiqing Zhang,&nbsp;Yibo Tang,&nbsp;Zixi Liu,&nbsp;Zhangyong Hong,&nbsp;Jian Zhang","doi":"10.1021/cbmi.5c00265","DOIUrl":"10.1021/cbmi.5c00265","url":null,"abstract":"<p>Adjuvants can stimulate the innateimmune system independentlyof vaccine antigens. Elucidating their spatiotemporal distributionacross biological scales and correlating this distribution with immuneactivation are of considerable significance. A cross-scale photoacoustic/fluorescencedual-modality imaging system was developed to enable dynamic trackingof the adjuvant from the cellular to the organismal level. Single-cellimaging results revealed that macrophages exhibited differential activationafter the uptake of different adjuvants. Notably, high-resolutionphotoacoustic microscopy (PAM) revealed size-dependent differencesin the intravascular kinetics of adjuvant in zebrafish tail veinsas well as their accumulation and clearance during circulation. Ina tail amputation-induced inflammation model, fluorescence microscopydemonstrated that CpG oligodeoxynucleotides elicited a 5-fold enhancementin immune cell recruitment efficiency compared to controls, whilenano/micron aluminum adjuvants induced 2-fold and 1.5-fold increases,respectively. The dual-modality photoacoustic/fluorescence systemquantitatively established that these immune outcomes directly originatefrom the spatiotemporal dynamics of the adjuvant. By bridging single-cellphagocytic behavior with systemic biodistribution, this study deciphereda causal mechanism. The activation of the immune system by adjuvantsresults from the synergistic interaction of their chemical compositionsand spatiotemporal distribution. This imaging-driven framework offersquantitative design principles for precision vaccine adjuvants.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":"4 7","pages":"1464–1476"},"PeriodicalIF":6.7,"publicationDate":"2026-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147877729","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}