Journal of Neuroscience Methods最新文献

{"title":"Detection of subarachnoid hemorrhage by bilateral transcranial laser doppler fluxmetry allows long-term studies in mice","authors":"Biyan Nathanael Harapan ,&nbsp;Julian Schwarting ,&nbsp;Andrea Cattaneo ,&nbsp;Dilara Ozsavas ,&nbsp;Uta Mamrak ,&nbsp;Gian Marco Calandra ,&nbsp;Xiangjiang Lin ,&nbsp;Nikolaus Plesnila ,&nbsp;Nicole Angela Terpolilli","doi":"10.1016/j.jneumeth.2025.110625","DOIUrl":"10.1016/j.jneumeth.2025.110625","url":null,"abstract":"<div><h3>Background</h3><div>The murine middle cerebral artery (MCA) perforation model is widely used to study subarachnoid hemorrhage (SAH) but typically requires invasive intracranial pressure (ICP) monitoring to prove successful induction of hemorrhage. However, ICP probe placement causes substantial additional parenchymal damage, confounding long-term structural and functional assessments. Therefore, we investigated whether bilateral transcranial cerebral blood flow (CBF) monitoring, which does not cause any parenchymal damage, can reliably detect SAH induction in mice.</div></div><div><h3>New method</h3><div>SAH was induced in 25 male C57BL/6 mice with simultaneous ICP and bilateral CBF monitoring over the MCA territory. An additional 40 mice underwent SAH surgery using only bilateral CBF monitoring (without ICP monitoring). Ipsilateral and contralateral CBF and ICP curves were analyzed to identify reliable predictors of successful SAH induction.</div></div><div><h3>Results</h3><div>MR imaging revealed progressive parenchymal injury caused by the ICP probe. SAH was reproducibly induced without ICP monitoring when bilateral CBF dropped &gt; 50 % from baseline within 90 s after perforation. Using this minimally invasive approach, SAH could be consistently induced by different surgeons, with severity and outcomes comparable to those with ICP monitoring.</div></div><div><h3>Comparison with existing methods and conclusion</h3><div>ICP probe placement introduces parenchymal damage and may therefore limit the validity of long-term outcome studies. Our findings suggest that bilateral CBF monitoring can effectively replace ICP measurement, reducing invasiveness and enabling valid long-term SAH experiments in mice.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"426 ","pages":"Article 110625"},"PeriodicalIF":2.3,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145534643","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Uncertainty in population receptive field estimates revealed by variational qPRF","authors":"Sebastian Waz ,&nbsp;Yalin Wang ,&nbsp;Zhong-Lin Lu","doi":"10.1016/j.jneumeth.2025.110628","DOIUrl":"10.1016/j.jneumeth.2025.110628","url":null,"abstract":"<div><h3>Background:</h3><div>Population receptive field (pRF) modeling is a cornerstone of retinotopic mapping in visual neuroscience, enabling precise mapping of visual stimulus processing in the human brain. However, pRF estimates are influenced by multiple sources of variability, including scanner properties, neurovascular coupling, physiological noise, and task-related factors. Traditionally, these estimates are treated as definitive because quantifying variance has been computationally infeasible.</div></div><div><h3>New method:</h3><div>We introduce qPRF-v, an innovative extension of the qPRF software which computes pRF point estimates over 1,000 times faster than existing packages while preserving goodness-of-fit (Waz et al., 2025a).</div></div><div><h3>Results:</h3><div>Leveraging variational inference, qPRF-v efficiently approximates the posterior variance of pRF model parameters, revealing substantial uncertainty, particularly in parameters governing neural dynamics (e.g., compressive exponent) compared to those defining receptive field centers.</div></div><div><h3>Comparison with existing methods:</h3><div>To evaluate the impact of variance quantification, we compared population-level retinotopic maps generated via simple averaging (the prior standard) and <span><math><msup><mrow><mi>R</mi></mrow><mrow><mn>2</mn></mrow></msup></math></span> weighted averaging with those using inverse-variance weighted averaging. Inverse-variance weighted averaging significantly enhanced map quality, yielding naturally denoised representations with improved eccentricity detail.</div></div><div><h3>Conclusions:</h3><div>By directly addressing variability, qPRF-v not only increases the accuracy of retinotopic mapping but also enables more robust, individualized analyses of visual processing. This advancement has profound implications for studying visual function in health and disease, paving the way for personalized neuroscience and improved brain atlases for research and clinical applications.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"426 ","pages":"Article 110628"},"PeriodicalIF":2.3,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145570311","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"High-density multielectrode array (MEA) recordings in a rodent glioma model","authors":"Ferdinand Forberger ,&nbsp;Fabiana Santana Kragelund ,&nbsp;Katrin Porath ,&nbsp;Rüdiger Köhling ,&nbsp;Falko Lange ,&nbsp;Timo Kirschstein","doi":"10.1016/j.jneumeth.2025.110623","DOIUrl":"10.1016/j.jneumeth.2025.110623","url":null,"abstract":"<div><h3>Background</h3><div>High-grade gliomas are aggressive primary brain tumors that affect the surrounding brain tissue, which can lead to neurological deficits, epilepsy, cognitive deficits, and tumor progression. Research has focused on glutamatergic alterations, but the understanding of the underlying network mechanisms is still scarce.</div></div><div><h3>New method</h3><div>To investigate how gliomas might affect neural network integrity and capacity for spiking activity, we induced network bursting events in neocortical slices from F98 glioma-bearing rats. Network activity was recorded using a 64 × 64 high-density multielectrode array to assess the network's capacity for extended periods of high activity. We describe glioma cell implantation, brain slice preparation, and detailed data analysis of electrophysiological recordings, focusing on computational methods for characterization of the network-wide bursting events.</div></div><div><h3>Results</h3><div>Spike sorting was performed using Kilosort4 and employed to identify network bursts. Burst origins were estimated from multi-unit activity, and network burst propagation speed was assessed. Network-level functional connectivity was analyzed using metrics derived from graphs based on the spike time tiling coefficient. Our results indicate reduced network integrity in glioma-bearing slices.</div></div><div><h3>Comparison with existing methods</h3><div>The combination of a high-density multielectrode array with Kilosort4 is advantageous over conventional recording techniques in terms of detecting network bursting and plotting center-of-activity trajectories.</div></div><div><h3>Conclusions</h3><div>This protocol provides a robust platform for dissecting network activity and establishes a basis for future mechanistic studies. We also discuss the limitations and experimental challenges of our preclinical approach to investigating glioma-associated effects on surrounding neuronal networks.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"426 ","pages":"Article 110623"},"PeriodicalIF":2.3,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145495893","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparing human annotation and machine learning models for optimizing zebrafish behavioral classification in seizure analysis","authors":"Barbara D. Fontana ,&nbsp;Camilla W. Pretzel ,&nbsp;Mariana L. Müller ,&nbsp;Kimberly Fontoura ,&nbsp;Khadija A. Mohammed ,&nbsp;Eduarda T. Saccol ,&nbsp;Falco L. Gonçalves ,&nbsp;Angela E. Uchoa ,&nbsp;Carolina C. Jardim ,&nbsp;Isabella P. Silva ,&nbsp;Rossano M. Silva ,&nbsp;Hevelyn S. Moraes ,&nbsp;Cássio M. Resmim ,&nbsp;Julia Canzian ,&nbsp;Denis B. Rosemberg","doi":"10.1016/j.jneumeth.2025.110603","DOIUrl":"10.1016/j.jneumeth.2025.110603","url":null,"abstract":"<div><h3>Background</h3><div>Accurate and scalable behavioral annotation remains a challenge in behavioral neuroscience. Manual scoring is time-consuming, variable across annotators, and may overlook transient behaviors critical for phenotyping. By learning from annotated datasets, supervised machine learning (ML) enables automated classification of behavior with high consistency and reduced bias.</div></div><div><h3>New method</h3><div>We benchmarked five supervised ML algorithms, Random Forest, XGBoost, Support Vector Machine, k-Nearest Neighbors, and Multilayer Perceptron (MLP), and compared data against expert human annotations of seizure-like behaviors in adult zebrafish. Twelve trained raters annotated over 43,000 video frames, enabling direct comparison of model performance with human annotation. After frame-level analysis, we also applied behavior-informed filters and then evaluated block-level temporal aggregation.</div></div><div><h3>Results</h3><div>Annotation variability was driven by behavioral complexity, with ambiguous behaviors showing the lowest agreement. Random Forest, XGBoost, and MLP achieved the highest accuracy and post-processing based on posture and velocity improved classification by filtering false positives. Block-level aggregation enhanced accuracy through temporal smoothing but masked short-lived behaviors critical for detecting subtle phenotypes.</div></div><div><h3>Comparison with existing methods</h3><div>Most zebrafish seizure studies rely on manual scoring or single-model ML applications. Direct comparisons between multiple ML algorithms and human annotations are rare. Our study uniquely integrates large-scale manual scoring with model benchmarking and temporal resolution strategies, offering insight into reproducibility and scalability in behavioral phenotyping.</div></div><div><h3>Conclusions</h3><div>This study advances automated behavioral analysis in zebrafish by demonstrating the strengths and limitations of machine learning compared to human annotation, and emphasizes how choices in temporal resolution and behavioral classification influence reproducibility and interpretability.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"425 ","pages":"Article 110603"},"PeriodicalIF":2.3,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145355088","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"RTFED, an open-source versatile tool for home-cage monitoring of behaviour and fibre photometry recording in mice","authors":"Hamid Taghipourbibalan,&nbsp;James Edgar McCutcheon","doi":"10.1016/j.jneumeth.2025.110604","DOIUrl":"10.1016/j.jneumeth.2025.110604","url":null,"abstract":"<div><h3>Background</h3><div>Conventional approaches for studying feeding and reward-driven behaviours require frequent animal handling or relocation of animals to specialized chambers, inducing stress, confounding behavioural outcomes, and limiting continuous (24/7) data collection. In recent years, the Feeding Experimentation Device (FED3) has emerged as a major advance, offering programmable modes of operation, affordable costs, and flexibility for investigating a range of feeding and operant behaviours. However, certain limitations prevent researchers from fully harnessing the FED3’s capabilities in a user-friendly manner.</div></div><div><h3>New method</h3><div>Here, we present the Real-time and Remote FED3 (RTFED) developed for continuous and online home-cage monitoring of mice, video recording behaviours and fibre photometry recording.</div></div><div><h3>Results</h3><div>Validation experiments confirm RTFED integrates well with FED3 to log and transmit behavioural events in real-time. It also incorporates event-triggered video capture through USB cameras, providing additional observational depth. Moreover, RTFED handles TTL signals to the fibre photometry system allowing precise behaviour-neural synchronization.</div></div><div><h3>Comparison with existing methods</h3><div>A key strength of RTFED is its easily customizable architecture, enabling researchers to tailor both software and hardware configurations to meet specific experimental objectives. This flexibility, together with features such as remote data logging and email notifications that allow timely adjustments and animal welfare monitoring based on behavioural observations, substantially reduces animal disturbance and researcher intervention and labour.</div></div><div><h3>Conclusions</h3><div>By offering a cost-effective and modifiable alternative to proprietary commercial solutions, RTFED broadens accessibility, heightens reproducibility, and deepens investigations into feeding and reward-driven behaviours in home-cage settings, ultimately improving the quality and translational relevance of behavioural research.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"425 ","pages":"Article 110604"},"PeriodicalIF":2.3,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145409297","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development and evaluation of a 3D-engineered neural co-culture system: Impacts on oxidative stress, pentose phosphate pathway, trace element and mineral metabolisms","authors":"Duygu Aydemir ,&nbsp;Buse Keleş ,&nbsp;İrem Polat ,&nbsp;Ecem Metin ,&nbsp;Emel Sokullu ,&nbsp;Nuriye Nuray Ulusu","doi":"10.1016/j.jneumeth.2025.110614","DOIUrl":"10.1016/j.jneumeth.2025.110614","url":null,"abstract":"<div><h3>Background</h3><div>Co-culturing multiple cell types within three-dimensional (3D) systems enhances the capacity to investigate intricate cell-cell and cell-microenvironment interactions, offering deeper insights into multicellular dynamics. This study comprehensively investigates the role of cellular metabolism within 3D cell culture models, offering a detailed examination of the underlying metabolic processes.</div></div><div><h3>New method</h3><div>In this study, a three-dimensional co-culture system was developed by encapsulating human neuroblastoma (SH-SY5Y) and human umbilical vein endothelial (HUVEC) cells within photopolymerized gelatin methacrylate (GelMA) hydrogels using photomasks for studying multiplex neural co-cultures. The structural characteristics of the hydrogel were analyzed using Fourier-transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). Cell proliferation and antioxidant enzyme activities, including glucose 6-phosphate dehydrogenase (G6PD), 6-phosphoglucanate dehydrogenase (6-PGD), glutathione reductase (GR), glutathione s-transferase (GST), and glutathione peroxidase (GPx) were measured. The levels of trace elements and minerals were also quantified.</div></div><div><h3>Results</h3><div>The 3D-co-culture system can be considered non-toxic based on ISO 10993–5 since cell viability did not reduce below 80 % on the 7th day compared to day 0. The 3D model did not adversely affect the indicated enzymes in the co-culture system for up to 7 days. Na, Ca, Cu, Zn, and Mg levels significantly increased in the first, 4^th, and 7th days compared to day 0.</div></div><div><h3>Comparison with existing methods</h3><div>Although photomask-based patterning of GelMA scaffolds has been previously demonstrated, our approach is unique as it combines multiplex photomask fabrication with the co-culture of neuronal and endothelial cells. Additionally, we measure multiple metabolic pathways, including the pentose phosphate pathway (PPP) and antioxidant enzymes, as well as the dynamics of trace and mineral elements in spatially defined neurovascular co-cultures. This integration allows for the simultaneous production of numerous geometrically controlled replicates and provides the first comprehensive assessment of PPP activity alongside trace element dynamics in engineered neural co-cultures.</div></div><div><h3>Conclusion</h3><div>This study highlights the benefits of using GelMA-based constructs in supporting viable and metabolically active cells in a 3D environment and presents a robust framework of methodologies that can be employed in future research to elucidate the complex metabolic dynamics in 3D environments, in tissue engineering, disease modeling, and drug development.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"425 ","pages":"Article 110614"},"PeriodicalIF":2.3,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145426652","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Detection of eye movements and eye blinks using a portable two-channel EEG platform","authors":"Xingyi Zhong ,&nbsp;Guangye Li ,&nbsp;Ce Xu ,&nbsp;Ruijie Luo ,&nbsp;Jianjun Meng ,&nbsp;Gerwin Schalk","doi":"10.1016/j.jneumeth.2025.110616","DOIUrl":"10.1016/j.jneumeth.2025.110616","url":null,"abstract":"<div><h3>Background:</h3><div>The ability to detect eye movements can facilitate human–computer interaction (HCI) and may complement brain–computer interfaces (BCIs). Recent studies have shown that multi-channel EEG systems can provide information about eye movements, but these systems can be bulky and/or require complex setup.</div></div><div><h3>New method:</h3><div>We introduce a portable, two-channel EEG platform that can be placed in seconds and detect eye blinks/movements and gaze trajectories. Forty adults performed cued blinks and horizontal/vertical gaze shifts; 21 EEG features were extracted, and machine learning models were evaluated with leave-one-subject-out validation.</div></div><div><h3>Results:</h3><div>Our system effectively identified eye blinks (avg. detection accuracy of 95%, 50% chance) and horizontal eye movements (avg. accuracy of 94%, 33% chance), and showed decreased performance detecting vertical eye movements (avg. accuracy of 60%, 33% chance). It was also able to predict horizontal and vertical eye movement trajectories (r = 0.79 and r = 0.14, respectively).</div><div><strong>Comparison with existing methods:</strong> Classification accuracies for eye blinks and horizontal eye movements using our system with only two electrodes are comparable to those previously reported only for complex multi-channel EEG/EOG setups.</div></div><div><h3>Conclusion:</h3><div>This study provides evidence, for the first time, that a wearable EEG device can give substantial information about eye blinks and eye movements. With further refinements, this approach may enable portable solutions for real-world HCI and BCI applications.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"425 ","pages":"Article 110616"},"PeriodicalIF":2.3,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145431731","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The effects of MEP counts on intra- and intersession reliability of single-pulse TMS-induced MEPs in the wrist extensor muscle of healthy individuals","authors":"Hamed Alanazi ,&nbsp;Prue Morgan ,&nbsp;Shapour Jaberzadeh","doi":"10.1016/j.jneumeth.2025.110621","DOIUrl":"10.1016/j.jneumeth.2025.110621","url":null,"abstract":"<div><h3>Background</h3><div>Motor evoked potentials (MEPs) exhibit high intra-individual variability, which can compromise reliability. One suggested approach to reduce this variability is averaging multiple MEPs. However, there is no evidence regarding the optimal number of MEPs required to obtain the most reliable amplitude and latency measurements from the extensor carpi radialis (ECR) muscle.</div></div><div><h3>New method</h3><div>Healthy participants attended two sessions. In the first session, two sets of TMS-induced MEPs were recorded with a 20-minute interval to evaluate intra-session reliability. The second session was conducted after at least 48 h to assess inter-session reliability. Reliability was quantified using intraclass correlation coefficients (ICCs), one-way ANOVA, and Bland–Altman analysis.</div></div><div><h3>Results</h3><div>For MEP amplitude, both intra- and inter-session ICC values were excellent across all MEP block sizes, with reliability improving as the number of MEPs per block increased. For MEP latency, 10 MEPs were sufficient to achieve excellent intra-session reliability, whereas inter-session reliability ranged from good to excellent, with larger blocks (16–25 MEPs) providing the highest ICC values.</div></div><div><h3>Comparison with existing methods</h3><div>This study is the first to examine MEP reliability in the ECR muscle. Compared with previous methods, including monophasic TMS in other muscles, our protocol shows that MEP latency in the ECR requires larger MEP blocks to achieve high inter-session reliability.</div></div><div><h3>Conclusions</h3><div>This study suggests that averaging MEPs from the ECR yields highly reproducible amplitude measurements with few blocks, whereas MEP latency requires more blocks to achieve high inter-session reliability.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"425 ","pages":"Article 110621"},"PeriodicalIF":2.3,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145505058","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A modified α-synuclein seed amplification assay in Lewy body dementia using Raman spectroscopy and machine learning analysis","authors":"Nathan P. Coles ,&nbsp;Suzan Elsheikh ,&nbsp;Alaa Gouda ,&nbsp;Agathe Quesnel ,&nbsp;Lucy Butler ,&nbsp;Ojodomo J. Achadu ,&nbsp;Meez Islam ,&nbsp;Karunakaran Kalesh ,&nbsp;Annalisa Occhipinti ,&nbsp;Claudio Angione ,&nbsp;Jon Marles-Wright ,&nbsp;David J. Koss ,&nbsp;Alan J. Thomas ,&nbsp;Tiago F. Outeiro ,&nbsp;Panagiota S. Filippou ,&nbsp;Ahmad A. Khundakar","doi":"10.1016/j.jneumeth.2025.110617","DOIUrl":"10.1016/j.jneumeth.2025.110617","url":null,"abstract":"<div><h3>Background</h3><div>Lewy body dementias (LBD), comprising dementia with Lewy bodies (DLB) and Parkinson’s disease dementia (PDD), are defined by misfolded α-synuclein aggregation. Seed amplification assays (SAAs), such as RT-QuIC, enable sensitive detection of α-synuclein aggregates but typically provide binary readouts and require fluorescence labeling. Raman spectroscopy offers a label-free approach to detect subtle biochemical changes, and its diagnostic potential can be enhanced with machine learning.</div></div><div><h3>Objectives</h3><div>This proof-of-concept study aimed to evaluate whether Raman spectroscopy combined with machine learning can improve SAA-based discrimination of LBD from controls in cerebrospinal fluid (CSF).</div></div><div><h3>Methods</h3><div>We analyzed a small number of post-mortem CSF samples from pathologically confirmed DLB (n = 2), PDD (n = 2), and controls (n = 2) using a 7-day SAA. Raman spectra were collected on Days 1, 4, and 7 and analyzed using principal component analysis (PCA) and uniform manifold approximation and projection (UMAP).</div></div><div><h3>Results</h3><div>Following SAA, both PCA and UMAP distinguished combined LBD samples from controls within 24 h (Day 1), reflecting biochemical changes consistent with α-synuclein fibrillation. Spectral shifts indicated decreased α-helical content with increased β-sheet structures. No consistent separation between DLB and PDD was observed.</div></div><div><h3>Conclusion</h3><div>This preliminary study demonstrates that combining Raman spectroscopy with machine learning can enable rapid, label-free detection of disease-specific changes. Despite the very limited sample size, these findings highlight the potential of this novel workflow and strongly warrant its validation in larger cohorts.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"425 ","pages":"Article 110617"},"PeriodicalIF":2.3,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145517762","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Impact of tissue storage time on immunodetection of c-Fos and GAD67 in the rat brain","authors":"Stoyan Dimitrov ,&nbsp;Xia Shan ,&nbsp;Jan Born ,&nbsp;Marion Inostroza","doi":"10.1016/j.jneumeth.2025.110602","DOIUrl":"10.1016/j.jneumeth.2025.110602","url":null,"abstract":"<div><h3>Background</h3><div>Activity-dependent markers such as c-Fos, a rapid indicator of neuronal activation, and GAD67, an enzyme essential for GABA synthesis in inhibitory neurons, are extensively employed to elucidate neural circuit dynamics. Given that many studies span extended periods with multiple experimental groups, it is crucial to ensure long-term storage of non-frozen brain tissue does not compromise immunodetection.</div></div><div><h3>New method</h3><div>Here, we evaluated the impact of storage duration on the immunodetection of c-Fos and GAD67 in rat brains. Intact brains, fixed in paraformaldehyde, were stored at 4 °C in phosphate-buffered saline with sodium azide to prevent bacterial growth. Brains were assessed at two storage durations - short (1.5 months) and prolonged (10 months). Brain sections were immunostained for c-Fos and GAD67 and imaged by confocal microscopy.</div></div><div><h3>Results</h3><div>We observed robust c-Fos immunoreactivity across multiple regions of the medial prefrontal cortex, hippocampus, and neocortex, with no significant differences attributable to storage duration. Additionally, quantifications of GAD67-positive cells and cells co-labeled for c-Fos/ GAD67 confirmed that immunodetection of inhibitory neurons remains intact when whole brains are stored for up to 10 months. In contrast, prolonged storage of brain slices strongly reduced c-Fos, but increased GAD67 staining.</div></div><div><h3>Comparison with existing method(s)</h3><div>The stability of c-Fos and GAD67 in tissue stored long-term at 4 °C remains untested.</div></div><div><h3>Conclusions</h3><div>These findings underscore that whereas intact brains can be safely stored for prolonged periods at 4°C without compromising antigenicity, brain slices are highly susceptible to storage-induced deterioration - insights important for planning and interpreting immunohistochemical studies in neuroscience.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"425 ","pages":"Article 110602"},"PeriodicalIF":2.3,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145344947","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}