Journal of Neuroscience Methods最新文献

{"title":"Action tremor kinematics exhibit features of stabilization dysfunction in essential tremor patients","authors":"Madelyn Mendlen ,&nbsp;Rex N. Tien ,&nbsp;Keanu Chee ,&nbsp;Jonathan P. Platt ,&nbsp;Jannika Machnik ,&nbsp;Eric Bayman ,&nbsp;Chiagoziem Anigbogu ,&nbsp;Drew Kern ,&nbsp;Steven G. Ojemann ,&nbsp;John A. Thompson ,&nbsp;Daniel R. Kramer","doi":"10.1016/j.jneumeth.2026.110680","DOIUrl":"10.1016/j.jneumeth.2026.110680","url":null,"abstract":"<div><h3>Background</h3><div>Action tremor that begins with voluntary movement is a common manifestation of essential tremor (ET). We investigated a novel approach to detect reach direction and tremor direction across naturalistic reaches, elucidating this relationship.</div></div><div><h3>Methods</h3><div>ET patients undergoing awake DBS implantation surgery performed a center-out reaching task, which elicited multi-joint, naturalistic reaching movements made to evenly spaced radial targets. This project utilized a deep-learning based markerless motion tracking system to monitor movements in 3-dimensions. For analysis, to test novel hypotheses, we compared the direction of the tremor vectors (during hold) to the direction of the reach vector, as well as to the other tremor direction vectors for each subject.</div></div><div><h3>Results</h3><div>We analyzed 13 limb recordings from 9 subjects. Tremor directions were not significantly dependent on preceding reach directions in any recording (p &gt; 0.05). The distribution of observed tremor directions was significantly different from the expected distribution of random vectors for all 11 analyzed recordings (likelihood of observed tremor directions vs. randomly generated vectors, p &lt; 0.01).</div></div><div><h3>Comparison with existing methods</h3><div>Our markerless capture method for tremor measurement does not rely on accelerometry or electromyography and does not require fiducials for tracking and combines two open-source python toolkits to leverage computer vision for three-dimensional pose estimation. Further we test a novel solution for measuring the relationship between tremor direction and movement.</div></div><div><h3>Conclusions</h3><div>Here, we describe a novel method of tremor detection. Our findings suggest that an action tremor is not generated as an oscillation of a function involved in reach, such as braking, but is consistent with a function involved in stabilization to maintain a posture.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"428 ","pages":"Article 110680"},"PeriodicalIF":2.3,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145989690","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":"Image segmentation and registration of carp brain tissue slices oriented to brain atlas construction","authors":"Yanhong Yan ,&nbsp;Yang Zhao ,&nbsp;Yong Peng ,&nbsp;Lingjun Han ,&nbsp;Shuhao Sun ,&nbsp;Yudong Wen ,&nbsp;Xueying Dong","doi":"10.1016/j.jneumeth.2025.110670","DOIUrl":"10.1016/j.jneumeth.2025.110670","url":null,"abstract":"<div><h3>Background</h3><div>Brain atlas is an important tool to provide information on the location and function of brain motor nuclei, and the segmentation and registration of brain tissue slice images is the necessary basis for building brain atlases.</div></div><div><h3>New method</h3><div>In order to construct the carp brain atlas, the segmentation and the registration of carp brain tissue images were studied in this study. In this study, carp brain tissue sections were prepared by the method of pre-fixation of brain tissue specimens and paraffin tissue sections with HE staining techniques, and a multi-threshold image segmentation algorithm based on HSI color space was selected for image segmentation according to the image characteristics of carp brain tissue sections. In the aspect of image registration, a method of selecting feature points for registration based on the morphological and structural characteristics of different brain regions of carp was used for image registration of brain tissue slices.</div></div><div><h3>Results</h3><div>The results showed that the image segmentation and registration algorithm used in this study can meet the experimental requirements and conform to the structural characteristics of carp brain in spatial position.</div></div><div><h3>Comparison with existing methods</h3><div>The traditional brain mapping is drawn manually by humans, which is time-consuming and labor-intensive, and is limited to two dimensions. Image segmentation technology can automatically identify the contours of brain regions, saving manpower and promoting three-dimensional reconstruction and digital research.</div></div><div><h3>Conclusions</h3><div>This study laid a foundation for the construction of stereotaxic map of carp brain.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"428 ","pages":"Article 110670"},"PeriodicalIF":2.3,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145917662","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":"OpenFieldAI – new open-source AI based software for tracking rodents and training open field test models","authors":"Muzammil Kabier ,&nbsp;Shamili Mariya Varghese ,&nbsp;K.V. Athira ,&nbsp;Mohamed A. Abdelgawad ,&nbsp;Mohammed M. Ghoneim ,&nbsp;Hailah M. Almohaimeed ,&nbsp;Sunil Kumar ,&nbsp;K.P. Sreekumar ,&nbsp;Ashok R. Unni ,&nbsp;Bijo Mathew","doi":"10.1016/j.jneumeth.2025.110667","DOIUrl":"10.1016/j.jneumeth.2025.110667","url":null,"abstract":"<div><div>Open field test (OFT) is one of the widely used pre-clinical models for assessing the exploratory, locomotion and anxiety behavior of rodents. OFT parameters are often analyzed manually or by using automated systems. Although many softwares exist for OFT analysis, the steep learning curve and the cost of commercial software leads the researcher to opt for traditional approaches. Manual analysis is riddled with observer bias which can lead to ambiguity in behavior classification. This leads to the preference of automated system over manual observation in a scientific context. Herein we present <strong>OpenFieldAI</strong>, which is an open-source python-based software which has an intuitive graphical user interface (GUI) interface which makes it beginner friendly for researcher. The software utilizes YOLO algorithm for the detection and tracking of rodents in OFT apparatus. When pre-trained models are unable to infer sufficiently, the ability to train models according to user criteria can be helpful. Single/multiple pre-recorded videos (mp4) or live video with external web cam can be given as input to calculate the parameters like speed, distance, time spent in/out of region of interest (ROI) and entries/exits with the generation of box centroid graph, heat map and line path which are crucial information that give scientific insights into the rodents neurobehavior. The automatic detection of central/peripheral region by min/max calculation in 2D plane and manual drawing of ROI contribute to the ease in use and collection of intricate information. To validate the software, we compared the observation of 5 parameters – total distance, speed, entries into the central region and time spend in central and peripheral regions from the readings of ANY-maze (commercial software) using Pearson correlation coefficient. Correlation within all the three groups was found to be above 0.9 which indicates the reliability of the new software. Moreover, User’s guide has been provided for proper utilization of the tool. OpenFieldAI which can be downloaded and installed in Windows OS for free at: <span><span>https://sourceforge.net/projects/openfieldai/</span><svg><path></path></svg></span></div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"428 ","pages":"Article 110667"},"PeriodicalIF":2.3,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145895820","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":"Corrigendum to “Proper reference selection and re-referencing to mitigate bias in single pulse electrical stimulation data” [J. Neurosci. Methods. 419, (2025) 110461]","authors":"Harvey Huang ,&nbsp;Joshua A. Adkinson ,&nbsp;Michael A. Jensen ,&nbsp;Mohammed Hasen ,&nbsp;Isabel A. Danstrom ,&nbsp;Kelly R. Bijanki ,&nbsp;Nicholas M. Gregg ,&nbsp;Kai J. Miller ,&nbsp;Sameer A. Sheth ,&nbsp;Dora Hermes ,&nbsp;Eleonora Bartoli","doi":"10.1016/j.jneumeth.2026.110690","DOIUrl":"10.1016/j.jneumeth.2026.110690","url":null,"abstract":"","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"428 ","pages":"Article 110690"},"PeriodicalIF":2.3,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146010757","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":"Synergistic integration of clinical and multi-omics data for early MCI diagnosis using an attention-based graph fusion network","authors":"Shuang Yu ,&nbsp;Jing Zhao ,&nbsp;Jing Ouyang ,&nbsp;Xiaming Wang ,&nbsp;Peng Kou ,&nbsp;Keying Zhu ,&nbsp;Ping Liu","doi":"10.1016/j.jneumeth.2025.110664","DOIUrl":"10.1016/j.jneumeth.2025.110664","url":null,"abstract":"<div><h3>Background</h3><div>Mild cognitive impairment (MCI), a precursor to Alzheimer’s disease (AD), requires precise early diagnosis. Single-omics approaches often miss disease complexity, motivating integrative and interpretable solutions.</div></div><div><h3>New method</h3><div>We present the Attention-based Multimodal Graph Fusion Network (A-MGFN), which integrates clinical, genomic, epigenomic, and transcriptomic data via biologically curated features – Clinico-Genetic Risk Score (CGRS), Curated Epigenomic Signature (CES), and Differential Expression Signature (DES). Each modality is encoded by a modality-specific graph convolutional network to capture higher-order intra-modal interactions, and a downstream attention module adaptively weights modalities for fusion.</div></div><div><h3>Results</h3><div>On the ADNI cohort, A-MGFN achieved an AUC of 0.86 ± 0.03 and an F1-score of 0.88 ± 0.03. Ablation and attention-weight analyses confirmed multi-omics synergy, with CES providing the largest marginal performance gains.</div></div><div><h3>Comparison with existing methods</h3><div>A-MGFN outperformed traditional machine-learning baselines and Graph Convolutional Network (GCN) frameworks (MO-GCAN, AD-GCN), with 5–7 percentage-point gains in F1-score, attributable to attention-guided fusion rather than fixed or unified-graph schemes.</div></div><div><h3>Conclusions</h3><div>A-MGFN offers a robust and interpretable multi-omics framework for early MCI detection and provides insights into modality contributions that may inform clinical translation. Its design is extensible to other neurodegenerative disorders (e.g., Parkinson’s disease).</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"428 ","pages":"Article 110664"},"PeriodicalIF":2.3,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145900618","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":"Interactions between external and internal attention processes during working memory","authors":"András Puszta ,&nbsp;Rene Huster ,&nbsp;Anne-Kristin Solbakk ,&nbsp;Ingrid Funderud ,&nbsp;Tor Endestad ,&nbsp;Venke Arntsberg Grane","doi":"10.1016/j.jneumeth.2026.110687","DOIUrl":"10.1016/j.jneumeth.2026.110687","url":null,"abstract":"<div><h3>Background</h3><div>The interaction between internally and externally driven attention (IDA and EDA) during visual working memory tasks has been primarily investigated separately. There is growing evidence that EDA and IDA interact closely during working memory. However, it seems that previous studies, especially those utilizing EEG, did not modulate EDA and IDA within the same task. Thus, there is a paucity of information on how the two processes interact with each other on the neural level.</div></div><div><h3>New method</h3><div>We developed a novel paradigm, the PUMA Task, to investigate the interplay between IDA and EDA. This task allows for simultaneous manipulation of IDA and EDA by varying the number of items to be remembered and the complexity of the visual stimuli.</div></div><div><h3>Results</h3><div>EEG power spectra, eye-tracking measures (pupillometry and fixation disparity) and behavioral response times were sensitive to the interaction between IDA and EDA. Alpha-beta cross-frequency coupling was primarily influenced by EDA.</div></div><div><h3>Comparison with existing methods</h3><div>Previous studies often modulated either EDA or IDA, but not both within the same task. The PUMA task enables concurrent manipulation of both attentional processes, providing a more comprehensive understanding of their interaction.</div></div><div><h3>Conclusions</h3><div>The PUMA task is a valuable tool for investigating the interplay between IDA and EDA during visual working memory. Our findings suggest that IDA and EDA are distinct attentional processes that dynamically interact, affecting working memory performance and neural correlates. Importantly, our newly developed task can be employed to more selectively assess attentional subprocesses in neuropsychiatric and neurological disorders.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"428 ","pages":"Article 110687"},"PeriodicalIF":2.3,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145979401","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":"Long term direct visualization of the photothrombotic cortical infarction through the intact skull of anesthetized mice","authors":"Juliette Leclerc,&nbsp;Théotime Briar,&nbsp;Caroline Derouck,&nbsp;Cheima Mortier,&nbsp;Célia Duclos,&nbsp;Karelle Bénardais,&nbsp;Eric Verin,&nbsp;Jean-Paul Marie,&nbsp;Julien Chuquet","doi":"10.1016/j.jneumeth.2026.110683","DOIUrl":"10.1016/j.jneumeth.2026.110683","url":null,"abstract":"<div><h3>Background</h3><div>To assess post-stroke functional recovery and develop new treatments, numerous preclinical models have been developed, including the photothrombotic stroke model. This reproducible and simple method induces a targeted ischemic lesion in a chosen cortical area following intravenous rose bengal injection and controlled illumination with a 532 nm laser. However, identifying the infarct’s location and extent <em>in vivo</em> requires sophisticated, time-consuming, and/or expensive tools such as MRI or advanced optical imaging techniques. Thus, we introduce here a simple and low-tech method.</div></div><div><h3>New method</h3><div>Our direct method takes advantage of the long-lasting fluorescence of rose bengal remaining in the damaged cortex and detectable through the intact skull using the same 532 nm laser.</div></div><div><h3>Result</h3><div>At the lesion site, we observed an emission spot glowing through the skull for several weeks after stroke induction. <em>Ex vivo</em> immunohistochemical analysis showed that rose bengal fluorescence remains confined to the lesion, precisely delineating the infarct's boundaries.</div></div><div><h3>Comparison with existing methods</h3><div>This technique simplifies lesion localization and guides subsequent <em>in vivo</em> investigations, such as probe implantation, optogenetic fiber placement, or targeted tissue sampling in the perilesional cortex, where neuroplasticity and repair processes occur.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"428 ","pages":"Article 110683"},"PeriodicalIF":2.3,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145928155","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":"Optimizing stereotaxic injection strategy for AAV-mediated corticospinal tract tracing in mice","authors":"Wei Wang ,&nbsp;Tehan Zhang ,&nbsp;Shaolong Li ,&nbsp;Wenzhao Wang ,&nbsp;Quanhe Jin ,&nbsp;Chi Zhang ,&nbsp;Jie Liu ,&nbsp;Haijian Sun ,&nbsp;Shiqing Feng","doi":"10.1016/j.jneumeth.2026.110679","DOIUrl":"10.1016/j.jneumeth.2026.110679","url":null,"abstract":"<div><h3>Background</h3><div>The corticospinal tract (CST) is a major descending motor pathway essential for voluntary motor control. While adeno-associated virus-mediated anterograde tracing is widely used to label CST projections in mice, what the best stereotaxic injection coordinates and post-injection intervals remain unclear.</div></div><div><h3>New method</h3><div>Here, we systematically evaluated eight cortical injection strategies, differing in anterior-posterior (AP) and medial-lateral (ML) coordinates, the number of injection sites, and post-injection intervals. CST labeling was quantitatively assessed at cervical 2, cervical 5, thoracic 2, thoracic 6, and lumbar 2 spinal levels using transduced axon count (TAC), mean fluorescence intensity (MFI) and transduced area within the dorsal columns, normalized to the C2 segment.</div></div><div><h3>Results</h3><div>Across the tested AP/ML coordinates and single- versus multi-site injections, TAC and MFI were broadly comparable across cervical and thoracic levels, with the exception of reduced L2 labeling in Group IV (AP +0.70 mm). Furthermore, reducing the post-injection interval from four weeks to two weeks did not compromise labeling efficiency.</div></div><div><h3>Comparison with existing methods</h3><div>Conventional CST tracing typically requires multiple injections and ≥ 4-week intervals, increasing complexity and duration. Our optimized single-injection, 2-week protocol achieves comparable labeling fidelity while reducing procedural burden and improving reproducibility.</div></div><div><h3>Conclusion</h3><div>We suggest that effective CST labeling from L2 and rostral segments can be achieved with a single-point injection at AP coordinates between 0.0 and + 0.7 mm (ML fixed at 1.2 mm) or ML coordinates from + 0.7 to + 1.5 mm (AP fixed at 0.0 mm). These results establish a simplified, reproducible strategy for CST tracing.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"428 ","pages":"Article 110679"},"PeriodicalIF":2.3,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145944572","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":"Rapid invisible frequency tagging (RIFT) with a consumer monitor: A proof-of-concept","authors":"Olaf Dimigen,&nbsp;Ioana Badea,&nbsp;Iarina Simon,&nbsp;Mark M. Span","doi":"10.1016/j.jneumeth.2025.110660","DOIUrl":"10.1016/j.jneumeth.2025.110660","url":null,"abstract":"<div><h3>Background</h3><div>Rapid Invisible Frequency Tagging (RIFT) enables neural frequency tagging at rates above the flicker fusion threshold, eliciting steady-state responses to flicker that is almost imperceptible. While RIFT has proven valuable for studying visuospatial attention, it has so far relied on costly projector systems, typically in combination with magnetoencephalography (MEG). The recent emergence of high-speed organic light-emitting diode (OLED) monitors for consumers suggests that RIFT may also be feasible with much more accessible hardware.</div></div><div><h3>New method</h3><div>Here, we provide a proof-of-concept demonstrating successful RIFT using a consumer-grade 480 Hz OLED monitor in combination with electroencephalography (EEG). We also share practical recommendations for achieving precise stimulus timing at 480 Hz with minimal frame drops.</div></div><div><h3>Results</h3><div>In a central fixation task, participants viewed a tapered disc stimulus flickering either centrally or peripherally. Luminance was modulated sinusoidally at 60 Hz or 64 Hz, frequencies at which the flicker was barely visible. Photodiode recordings confirmed that the monitor delivered accurate frame timing with few dropped frames. Cross-coherence analysis between occipital EEG channels and a photodiode revealed robust, frequency-specific neural tagging responses for central stimuli at both frequencies. In comparison, weaker coherence was observed for 60 Hz peripheral flicker.</div></div><div><h3>Conclusions</h3><div>Our findings demonstrate that RIFT can be reliably implemented using affordable stimulation hardware, a low-density EEG montage, and a minimal processing pipeline. We hope that this lowers barriers to entry, facilitating broader use of RIFT in basic research and in applied settings where cost and portability matter.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"428 ","pages":"Article 110660"},"PeriodicalIF":2.3,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145794079","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":"Screening channelrhodopsins using robotic intracellular electrophysiology and single cell sequencing","authors":"Samuel Ehrlich ,&nbsp;Alexandra D. VandeLoo ,&nbsp;Mohamed Badawy ,&nbsp;Mercedes M. Gonzalez ,&nbsp;Max Stockslager ,&nbsp;Aimei Yang ,&nbsp;Sapna Sinha ,&nbsp;Shahar Bracha ,&nbsp;Demian Park ,&nbsp;Benjamin Magondu ,&nbsp;Bo Yang ,&nbsp;Edward S. Boyden ,&nbsp;Craig R. Forest","doi":"10.1016/j.jneumeth.2025.110663","DOIUrl":"10.1016/j.jneumeth.2025.110663","url":null,"abstract":"<div><h3>Background:</h3><div>Our ability to engineer opsins is limited by an incomplete understanding of how sequence variations influence function. The vastness of opsin sequence space makes systematic exploration difficult.</div></div><div><h3>New method:</h3><div>In recognition of the need for datasets linking opsin genetic sequence to function, we pursued a novel method for screening channelrhodopsins to obtain these datasets. In this method, we integrate advances in robotic intracellular electrophysiology (<u>P</u>atch) to measure optogenetic properties (<u>E</u>xcite), harvest individual cells of interest (<u>P</u>ick) and subsequently sequence them (<u>S</u>equence), thus tying sequence to function.</div></div><div><h3>Results:</h3><div>We used this method to sequence more than 50 cells with associated functional characterization. We further demonstrate the utility of this method with experiments on heterogeneous populations of known opsins and single point mutations of a known opsin. Of these point mutations, we found C160W ablates ChrimsonR’s response to light.</div></div><div><h3>Conclusion and comparison to existing methods:</h3><div>Compared to traditional manual patch clamp screening, which is labor-intensive and low-throughput, this approach enables more efficient, standardized, and scalable characterization of large opsin libraries. This method can enable opsin engineering with large datasets to increase our understanding of opsin sequence–function relationships.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"428 ","pages":"Article 110663"},"PeriodicalIF":2.3,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145800610","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}