Journal of Neuroscience Methods最新文献

{"title":"Positive reinforcement-based magnet training permits social housing in catheterized squirrel monkeys","authors":"Alexandra N. Johansen ,&nbsp;Hector M. Figueroa ,&nbsp;Jacquelin C. Hecker ,&nbsp;Jazmyne Z. Taylor ,&nbsp;Evan T. Shukan ,&nbsp;Hank P. Jedema ,&nbsp;Charles W. Bradberry","doi":"10.1016/j.jneumeth.2024.110313","DOIUrl":"10.1016/j.jneumeth.2024.110313","url":null,"abstract":"<div><h3>Background</h3><div>Non-human primates play a critical role in neuroscience research. Though they are social animals, laboratory study requirements can sometimes require single housing and thereby prevent social housing.</div></div><div><h3>New Method</h3><div>To eliminate single housing and promote well-being within our squirrel monkey colony, we used positive reinforcement training in combination with magnetic/mechanical clasps and custom jackets to permit pair housing of catheterized squirrel monkeys used in behavioral studies.</div></div><div><h3>Results</h3><div>Adult <em>Saimiri boliviensis boliviensis</em> monkeys (<em>n</em> = 7) readily progressed through a six-stage training procedure for cooperative handling and transport from the home cage to the experimental testing rooms.</div><div>Comparison with existing methods and conclusions: Given the evidence of isolation induced stress and neurobiological consequences in multiple species, and consistent with an increased regulatory emphasis on social housing of non-human primates, the methods presented herein provide a method for handling squirrel monkeys in behavioral studies that is compatible with social housing.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"413 ","pages":"Article 110313"},"PeriodicalIF":2.7,"publicationDate":"2024-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142564383","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":"Pupillometry is sensitive to speech masking during story listening: A commentary on the critical role of modeling temporal trends","authors":"Andreas Widmann ,&nbsp;Björn Herrmann ,&nbsp;Florian Scharf","doi":"10.1016/j.jneumeth.2024.110299","DOIUrl":"10.1016/j.jneumeth.2024.110299","url":null,"abstract":"<div><div>An increase in pupil size is an important index of listening effort, for example, when listening to speech masked by noise. Specifically, the pupil dilates as the signal-to-noise ratio decreases. A growing body of work aims to assess listening effort under naturalistic conditions using continuous speech, such as spoken stories. However, a recent study found that pupil size was sensitive to speech masking only when listening to sentences but not under naturalistic conditions when listening to stories. The pupil typically constricts with increasing time on task during an experimental block or session, and it may be necessary to account for this temporal trend in experimental design and data analysis in paradigms using longer, continuous stimuli. In the current work, we re-analyze the previously published pupil data, taking into account a problematic constraint of randomization and time-on-task, and use the data to outline methodological solutions for accounting for temporal trends in physiological data using linear mixed models. The results show that, in contrast to the previous work, pupil size is indeed sensitive to speech masking even during continuous story listening. Furthermore, accounting for the temporal trend allowed modeling the dynamic changes in the speech masking effect on pupil size over time as the continuous story unfolded. After demonstrating the importance of accounting for temporal trends in the analysis of empirical data, we provide simulations, methodological considerations, and user recommendations for the analysis of temporal trends in experimental data using linear mixed models.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"413 ","pages":"Article 110299"},"PeriodicalIF":2.7,"publicationDate":"2024-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142467906","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":"Quantifying population-level neural tuning functions using Ricker wavelets and the Bayesian bootstrap","authors":"Laura Ahumada ,&nbsp;Christian Panitz ,&nbsp;Caitlin M. Traiser ,&nbsp;Faith E. Gilbert ,&nbsp;Mingzhou Ding ,&nbsp;Andreas Keil","doi":"10.1016/j.jneumeth.2024.110303","DOIUrl":"10.1016/j.jneumeth.2024.110303","url":null,"abstract":"<div><h3>Background</h3><div>Experience changes visuo-cortical tuning. In humans, re-tuning has been studied during aversive generalization learning, in which the similarity of generalization stimuli (GSs) with a conditioned threat cue (CS+) is used to quantify tuning functions. Previous work utilized pre-defined tuning shapes (generalization and sharpening patterns). This approach may constrain the ways in which re-tuning can be characterized since the tuning patterns may not match the prototypical functions.</div></div><div><h3>New method</h3><div>The present study proposes a flexible and data-driven method for precisely quantifying changes in tuning based on the Ricker wavelet function and the Bayesian bootstrap. This method was applied to EEG and psychophysics data from an aversive generalization learning paradigm.</div></div><div><h3>Results</h3><div>The Ricker wavelet model fitted the steady-state visual event potentials (ssVEP), alpha-band power, and detection accuracy data well. A Morlet wavelet function was used for comparison and fit the data better in some situations, but was more challenging to interpret. The pattern of re-tuning in the EEG data, predicted by the Ricker model, resembled the shapes of the best fitting a-priori patterns.</div></div><div><h3>Comparison with existing methods</h3><div>Although the re-tuning shape modeled by the Ricker function resembled the pre-defined shapes, the Ricker approach led to greater Bayes factors and more interpretable results compared to a-priori models. The Ricker approach was more easily fit and led to more interpretable results than a Morlet wavelet model.</div></div><div><h3>Conclusion</h3><div>This work highlights the promise of the current method for capturing the precise nature of visuo-cortical tuning, unconstrained by the implementation of a-priori models.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"413 ","pages":"Article 110303"},"PeriodicalIF":2.7,"publicationDate":"2024-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142467907","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":"T1- and T2-weighted MRI signal and histology findings in suboptimally fixed human brains","authors":"Eve-Marie Frigon ,&nbsp;Philippe Pharand ,&nbsp;Amy Gérin-Lajoie ,&nbsp;Liana Guerra Sanches ,&nbsp;Denis Boire ,&nbsp;Mahsa Dadar ,&nbsp;Josefina Maranzano","doi":"10.1016/j.jneumeth.2024.110301","DOIUrl":"10.1016/j.jneumeth.2024.110301","url":null,"abstract":"<div><div>Neuroscientific research that requires brain tissue depends on brain banks that provide very small tissue samples fixed by immersion in neutral-buffered formalin (NBF), while anatomy laboratories could provide full brain specimens. However, these brains are generally fixed by perfusion of the full body with solutions other than NBF generally used by brain banks, such as an alcohol-formaldehyde solution (AFS) that is typically used for dissection and teaching. Therefore, fixation quality of these brains needs to be assessed to determine their usefulness in post-mortem investigations through magnetic resonance imaging (MRI) and histology, two common neuroimaging modalities. Here, we report the characteristics of five brains fixed by full body perfusion of an AFS from our Anatomy Laboratory suspected of being poorly fixed, given the altered signal seen on T1w MRI scans <em>in situ</em>. We describe 1- the characteristics of the donors; 2- the fixation procedures applied for each case; 3- the tissue contrast characteristics of the T1w and T2w images; 4- the macroscopic tissue quality after extraction of the brains; 5- the macroscopic arterial characteristics and presence or absence of blood clots; and 6- four histological stains of the areas that we suspected were poorly fixed. We conclude that multiple factors can affect the fixation quality of the brain. Nevertheless, cases in which brain fixation is suboptimal, consequently altering the T1w signal, still have T2w of adequate gray-matter to white-matter contrast and may also be used for histology stains with sufficient quality.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"412 ","pages":"Article 110301"},"PeriodicalIF":2.7,"publicationDate":"2024-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142467908","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhancing non-invasive pre-surgical evaluation through functional connectivity and graph theory in drug-resistant focal epilepsy","authors":"Silvana Pelle ,&nbsp;Anna Scarabello ,&nbsp;Lorenzo Ferri ,&nbsp;Giulia Ricci ,&nbsp;Francesca Bisulli ,&nbsp;Mauro Ursino","doi":"10.1016/j.jneumeth.2024.110300","DOIUrl":"10.1016/j.jneumeth.2024.110300","url":null,"abstract":"<div><h3>Background</h3><div>Epilepsy, characterized as a network disorder, involves widely distributed areas following seizure propagation from a limited onset zone. Accurate delineation of the epileptogenic zone (EZ) is crucial for successful surgery in drug-resistant focal epilepsy. While visual analysis of scalp electroencephalogram (EEG) primarily elucidates seizure spreading patterns, we employed brain connectivity techniques and graph theory principles during the pre-ictal to ictal transition to define the epileptogenic network.</div></div><div><h3>Method</h3><div>Cortical sources were reconstructed from 40-channel scalp EEG in five patients during pre-surgical evaluation for focal drug-resistant epilepsy. Temporal Granger connectivity was estimated ten seconds before seizure and at seizure onset. Results have been analyzed using some centrality indices taken from Graph theory (Outdegree, Hubness). A new lateralization index is proposed by taking into account the sum of the most relevant hubness values across left and right regions of interest.</div></div><div><h3>Results</h3><div>In three patients with positive surgical outcomes, analysis of the most relevant Hubness regions closely aligned with clinical hypotheses, demonstrating consistency in EZ lateralization and location. In one patient, the method provides unreliable results due to the abundant movement artifacts preceding the seizure. In a fifth patient with poor surgical outcome, the proposed method suggests a wider epileptic network compared with the clinically suspected EZ, providing intriguing new indications beyond those obtained with traditional electro-clinical analysis.</div></div><div><h3>Conclusions</h3><div>The proposed method could serve as an additional tool during pre-surgical non-invasive evaluation, complementing data obtained from EEG visual inspection. It represents a first step toward a more sophisticated analysis of seizure onset based on connectivity imbalances, electrical propagation, and graph theory principles.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"413 ","pages":"Article 110300"},"PeriodicalIF":2.7,"publicationDate":"2024-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142467905","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":"Modified rat pup cerebrospinal fluid collection method","authors":"Jiaojiao Wang,&nbsp;Zhifang Dong,&nbsp;Xiuyu Shi","doi":"10.1016/j.jneumeth.2024.110302","DOIUrl":"10.1016/j.jneumeth.2024.110302","url":null,"abstract":"<div><h3>Background</h3><div>Cerebrospinal fluid (CSF) reflects biochemical changes in the brain due to its direct contact with brain interstitial fluid, making it a valuable tool for diagnosing and monitoring disease progression and therapeutic effectiveness in clinical practice. However, collecting CSF in animal studies, particularly from small animals like rat pups or mice, poses significant challenges.</div></div><div><h3>New method</h3><div>After attempting various reported protocols, we encountered difficulties in consistently obtaining sufficient CSF from rat pups (P7-P42). Consequently, we modified these methods and developed a protocol with controllable and precise parameters for each step, enhancing reproducibility across different researchers.</div></div><div><h3>Results</h3><div>The newly developed method enables rapid, single-operator, and reproducible CSF extraction while ensuring high-quality (the absorbance of the “quality control solution” at 415 nm &lt; 0.05 AU, an indicator of oxyhemoglobin contamination for the collected CSF samples) and high-yield samples (33 ± 2.128 μL for P7 pups, 34.10 ± 2.747 μL for P8 pups, 36.67 ± 3.997 μL for P9 pups, 36.90 ± 1.946 μL for P10 pups, 35.11 ± 3.285 μL for P10 hypoxic-ischemic brain damage (HIBD) pups and 51.70 ± 5.256 μL for P42 pups, respectively).</div></div><div><h3>Comparison with existing methods</h3><div>Unlike existing methods of CSF extraction in rat pups, our protocol has reproducible capillary pipette pulling parameters, controllable CSF quality indexes, and can be operated by a single person with high yield in a short time.</div></div><div><h3>Conclusions</h3><div>This paper provides a step-by-step comparison and discussion of the CSF collection process, establishing a method that enables a single operator to collect CSF rapidly, consistently, sufficiently, and with controlled quality.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"412 ","pages":"Article 110302"},"PeriodicalIF":2.7,"publicationDate":"2024-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142445875","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":"Localization of neurons from extracellular footprints","authors":"Mélina Scopin ,&nbsp;Giulia L.B. Spampinato ,&nbsp;Olivier Marre ,&nbsp;Samuel Garcia ,&nbsp;Pierre Yger","doi":"10.1016/j.jneumeth.2024.110297","DOIUrl":"10.1016/j.jneumeth.2024.110297","url":null,"abstract":"<div><h3>Background:</h3><div>High density microelectrode arrays (HD-MEAs) are now widely used for both <em>in-vitro</em> and <em>in-vivo</em> recordings, as they allow spikes from hundreds of neurons to be recorded simultaneously. Since extracellular recordings do not allow visualization of the recorded neurons, algorithms are needed to estimate their physical positions, especially to track their movements when the are drifting away from recording devices.</div></div><div><h3>New Method:</h3><div>The objective of this study was to evaluate the performance of multiple algorithms for neuron localization solely from extracellular traces (MEA recordings), either artificial or obtained from mouse retina. The algorithms compared included center-of-mass, monopolar, and grid-based algorithms. The first method is a barycenter calculation. The second algorithm infers the position of the cell using triangulation with the assumption that the neuron behaves as a monopole. Finally, grid-based methods rely on comparing the recorded spike with a projection of spikes of hypothetical neurons with different positions.</div></div><div><h3>Results:</h3><div>The Grid-Based algorithm yielded the most satisfactory outcomes. The center-of-mass exhibited a minimal computational cost, yet its average localization was suboptimal. Monopolar algorithms gave cell localizations with an average error of less than <span><math><mrow><mn>10</mn><mspace></mspace><mi>μ</mi><mi>m</mi></mrow></math></span>, but they had considerable variability and a high computational cost. For the grid-based method, the variability was smaller, with satisfactory performance and low computational cost.</div></div><div><h3>Comparison with Existing Method(s):</h3><div>The accuracy of the different localization methods benchmarked in this article had not been properly tested with ground-truth recordings before.</div></div><div><h3>Conclusion:</h3><div>The objective of this article is to provide guidance to researchers on the selection of optimal methods for localizing neurons based on MEA recordings.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"412 ","pages":"Article 110297"},"PeriodicalIF":2.7,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142400422","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optogenetic estimation of synaptic connections in brain slices","authors":"Tetsuhiko Kashima ,&nbsp;Takuya Sasaki ,&nbsp;Yuji Ikegaya","doi":"10.1016/j.jneumeth.2024.110298","DOIUrl":"10.1016/j.jneumeth.2024.110298","url":null,"abstract":"<div><h3>Background</h3><div>Detection of synaptic connections is essential for understanding neural circuits. By using optogenetics, current injection, and glutamate uncaging to activate presynaptic cells and simultaneously recording the subsequent response of postsynaptic cells, the presence of synaptic connections can be confirmed. However, these methods present throughput challenges, such as the need for simultaneous multicellular patch-clamp recording and two-photon microscopy. These challenges lead to a trade-off between sacrificing resolution and experimental throughput.</div></div><div><h3>New method</h3><div>We adopted the laser, typically used for local field ablation, and combined this with post hoc analysis. We successfully approximated the synaptic connection probabilities using only an epi-fluorescence microscope and single-cell recordings.</div></div><div><h3>Results</h3><div>We sequentially stimulated the channelrhodopsin 2-expressing cells surrounding the recorded cell and approximated the synaptic connection probabilities. This probability value was comparable to that obtained from simultaneous multi-cell patch-clamp recordings, which included more than 600 pairs.</div></div><div><h3>Comparison with existing methods</h3><div>Our setup allows us to estimate connection probabilities within 100 s, outperforming existing methods. We successfully estimated synaptic connection probabilities using only the optical path typically used by an epi-fluorescence microscope and single-cell recordings. It may also be suitable for dendritic ablation experiments.</div></div><div><h3>Conclusions</h3><div>The proposed method simplifies the estimation of connection probabilities, which is expected to advance the study of neural circuits in conditions such as autism and schizophrenia where connection probabilities vary. Furthermore, this approach is applicable not only to local circuits but also to long-range connections, thus increasing experimental throughput.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"412 ","pages":"Article 110298"},"PeriodicalIF":2.7,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142372095","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of neuronal differentiation in human adipose-derived stromal cells: morphological, molecular, and ultrastructural insights","authors":"Xiaodong Yuan ,&nbsp;Wen Li ,&nbsp;Yi Yuan ,&nbsp;Xuhong Zhu ,&nbsp;Yan Meng ,&nbsp;Qi Wu ,&nbsp;Qi Yan ,&nbsp;Pingshu Zhang","doi":"10.1016/j.jneumeth.2024.110296","DOIUrl":"10.1016/j.jneumeth.2024.110296","url":null,"abstract":"<div><h3>Objective</h3><div>Adipose-derived stromal cells (ADSCs) have shown promise as a potential source of neural differentiation. In this study, we investigated the morphological, molecular and ultrastructural features of ADSCs during neuronal differentiation.</div></div><div><h3>Methods</h3><div>ADSCs were induced in vitro and their differentiation was examined at different time points. Immunocytochemical staining was performed to detect the expression of neuron-specific markers NSE and MAP-2. Immunofluorescence double labeling and Western blot detected the co-expression of presynaptic markers (CaMKII, SynCAM1, SYN) and postsynaptic markers (PSD-95, Synapsin I). Scanning electron microscopy (SEM) was performed to detect the synaptic structural features of differentiated neurons.</div></div><div><h3>Results</h3><div>ADSCs showed diverse morphological features during differentiation, gradually acquiring a neuron-like spindle shape and organized arrangement. The expression of neuron-specific markers and synaptic markers peaked at 5 h of induction. Scanning electron microscopy showed polygonal protrusions of ADSC-derived neurons, and transmission electron microscopy showed characteristic ultrastructures such as nidus, synaptic vesicle-like structures, and tight junctions.</div></div><div><h3>Conclusion</h3><div>Our findings suggest that ADSCs differentiated for 5 h have neuronal features, including morphological, molecular, and ultrastructural resemblance to neurons, as well as the formation of synaptic structures. These insights contribute to a better understanding of ADSC-based neuronal differentiation and pave the way for future applications in regenerative medicine and neurodegenerative diseases.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"412 ","pages":"Article 110296"},"PeriodicalIF":2.7,"publicationDate":"2024-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142365471","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 method for quantitative spatial analysis of immunolabeled fibers at regenerative electrode interfaces","authors":"Michael Rosario ,&nbsp;Jingyuan Zhang ,&nbsp;Muhammad Irfan Kaleem ,&nbsp;Nikhil Chandra ,&nbsp;Ying Yan ,&nbsp;Daniel Moran ,&nbsp;Matthew Wood ,&nbsp;Wilson Z. Ray ,&nbsp;Matthew MacEwan","doi":"10.1016/j.jneumeth.2024.110295","DOIUrl":"10.1016/j.jneumeth.2024.110295","url":null,"abstract":"<div><h3>Background</h3><div>Regenerative electrodes are being explored as robust peripheral nerve interfaces for neuro-prosthetic control and sensory feedback. Current designs differ in electrode number, spatial arrangement, and porosity which impacts the regeneration, activation, and spatial distribution of fibers at the device interface. Knowledge of sensory and motor fiber distributions are important in optimizing selective fiber activation and recording.</div></div><div><h3>New Method</h3><div>We use confocal microscopy and immunofluorescence methods to conduct spatial analysis of immunolabeled fibers across whole nerve cross sections.</div></div><div><h3>Results</h3><div>This protocol was implemented to characterize motor fiber distribution within 3 macro-sieve electrode regenerated (MSE), 3 silicone-conduit regenerated, and 3 unmanipulated control rodent sciatic nerves. Total motor fiber counts were 1485 [SD: +/- 50.11], 1899 [SD: +/- 359], and 5732 [SD: +/- 1410] for control, MSE, and conduit nerves respectively. MSE motor fiber distributions exhibited evidence of deviation from complete spatial randomness and evidence of dispersion and clustering tendencies at varying scales. Notably, MSE motor fibers exhibited clustering within the central portion of the cross section, whereas conduit regenerated motor fibers exhibited clustering along the periphery.</div></div><div><h3>Comparison with Existing Methods</h3><div>Prior exploration of fiber distributions at regenerative interfaces was limited to either quadrant-based density analysis of randomly sampled subregions or qualitative description. This method extends existing sample preparation and microscopy techniques to quantitatively assess immunolabeled fiber distributions within whole nerve cross-sections.</div></div><div><h3>Conclusions</h3><div>This approach is an effective way to examine the spatial organization of fiber subsets at regenerative electrode interfaces, enabling robust assessment of fiber distributions relative to electrode arrangement.</div></div>","PeriodicalId":16415,"journal":{"name":"Journal of Neuroscience Methods","volume":"412 ","pages":"Article 110295"},"PeriodicalIF":2.7,"publicationDate":"2024-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142348461","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}