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1,218 results for “vesicles”
Problems with nanoparticle tracking analysis (NTA) of urine extracellular vesicles (uEVs)
<p>Urinary extracellular vesicle (uEV) proteins may be used as specific markers of kidney damage in various pathophysiological conditions. The nanoparticle-tracking analysis (NTA) appears to be the most useful method for the analysis of uEVs due to its ability to analyze particles below 300 nm. The NTA method has been used to measure the size and concentration of uEVs and also allows for a deeper analysis of uEVs based on their protein composition using fluorescence measurements. However, despite much interest in the clinical application of uEVs, their analysis using the NTA method is poorly described and requires meticulous sample preparation, experimental adjustment of instrument settings, and above all, an understanding of the limitations of the method. We present the problems encountered during analysis with possible solutions: the choice of sample dilution, the method of the presentation and comparison of results, photobleaching, and the adjustment of instrument settings for a specific analysis.</p> <p> </p> <p>Figure 1. Expressions of specific markers CD63 in protein-standardized samples detected with Western blot analysis; anti-CD 63 (HPA010088, Sigma-Aldrich, Saint Louis, MO, USA, 1:1000); secondary antibodies conjugated to horseradish peroxidase (554021, BD Pharmingen (BD Biosciences, San Jose, CA, USA) 1:10000).</p> <p> </p> <p>Nanoparticle-Tracking Analysis of uEVs. A NanoSight NS300 instrument (Malvern Panalytical, Malvern, UK) was used to determine the concentrations and sizes of the uEVs in the samples. The total number of extracellular vesicles was measured during the continuous flow of samples delivered from a syringe pump.</p> <p>Figure 2. Determination of the size and concentration of uEVs: dilution factor—1:100; laser—405 nm.</p> <p>Figure 3. Effect of dilution on total number of particles per milliliter and size of uEVs in nanoparticle tracking analysis: sample dilutions—1:100, 1:500, and 1:1000; laser—488 nm.</p> <p>Figure 5. Fluorescence-based nanoparticle-tracking analysis of CD 63 expression in uEVs: without 500 nm long-pass filter; with 500 nm long-pass filter; comparison of sizes and concentrations of uEVs without and with 500 nm long-pass filter; dilution factor—1:100; laser—488 nm; anti-CD 63 (HPA010088, Sigma-Aldrich); secondary antibodies conjugated to Alexa Fluor 488 fluorescent dye (ab150073-500, Abcam, Cambridge, MA, USA).</p> <p>Figure 6. Fluorescence-based nanoparticle-tracking analysis of podocin expression in uEVs: without 500 nm long-pass filter; with 500 nm long-pass filter; comparison of sizes and concentrations of uEVs without and with 500 nm long-pass filter; dilution factor—1:100; laser—488 nm; anti-podocin (P0372, Sigma-Aldrich); secondary antibodies conjugated to Alexa Fluor 488 fluorescent dye (ab150073-500, Abcam, Cambridge, MA, USA).</p>
Quantification of Giant Unilamellar Vesicle Fusion Products by High-Throughput Image Analysis - Imaging Flow Citometry Data
<p>Imaging flow citometry (IFC) datasets analysed in "Quantification of Giant Unilamellar Vesicle Fusion Products by High-Throughput Image Analysis" (under revision).</p> <p>The folders contain acquisitions of giant unilamellar vesicles (GUVs) for lipid exchange and content exchange controls, with file naming convention DATE_SAMPLE_REPLICATE.rif, content exchange is indicated by CE samples in the 20230228_CE.zip folder, lipid exchange by LE samples in the 20221222_LE.zip folder. 24 samples per set are included, triplicates of isolated P1 (DOPE Af488 0.6% in LE; Dex-Af488 40 uM for CE), P2 (DOPE Cy50.6% in LE; Dex-Af647 10 uM for CE), NC (P1 + P2 1:1), PC (DOPE Af488 0.3% + DOPE Cy5 0.3 in LE; Dex-Af488 20 uM + Dex-Af647 5 uM for CE), and M samples numbered 1 to 4, prepared by mixing P1, P2 and PC in different ratios (M1= 1:1:1; M2= 1:1:0.5; M3= 1:1:0.1; M4= 1:1:0.05).</p> <p>Only .rif files are provided, they have to be elaborated via compensation and application of an analysis template using the Amnis IDEAS software. Compensation matrices for lipid exchange (20230217_LEcom.ctm) and content exchange (20230217_CEcomp.ctm) are included, as well as the analysis template (Lipid_exchange_analysis_6.2.ast). Gating in the latter may have to be adjusted to analyse LE and CE experiments.</p> <p>10000 objects in the GUV population or 50000 objects in total were acquired in each file. The files were elaborated in batch mode, outputting the statistic reports (Statistics report CE.txt for CE; Statistics report LE.txt for LE) that were elaborated using an R scirpt (included, IFC_analysis.R) </p>
Ergolide Mediates Anti-Cancer Effects on Metastatic Uveal Melanoma Cells and Modulates their Cellular and Extracellular Vesicle proteomes
<p>Underlying dataset and extended dataset of the results described in the article "Ergolide Mediates Anti-Cancer Effects on Metastatic Uveal Melanoma Cells and Modulates their Cellular and Extracellular Vesicle proteomes".</p>
Efficient embryoid-based method to improve generation of optic vesicles from human induced pluripotent stem cells data
<p>Animal models have provided many insights into ocular development and disease, but they remain suboptimal for understanding human oculogenesis. Eye development requires spatiotemporal gene expression patterns and disease phenotypes can differ significantly between humans and animal models, with patient-associated mutations causing embryonic lethality reported in some animal models. The emergence of human induced pluripotent stem cell (hiPSC) technology has provided a new resource for dissecting the complex nature of early eye morphogenesis through the generation of three-dimensional (3D) cellular models. By using patient-specific hiPSCs to generate <em>in vitro </em>optic vesicle-like models, we can enhance the understanding of early developmental eye disorders and provide a pre-clinical platform for disease modelling and therapeutics testing. A major challenge of <em>in vitro </em>optic vesicle generation is the low efficiency of differentiation in 3D cultures. To address this, we adapted a previously published protocol of retinal organoid differentiation to improve embryoid body formation using a microwell plate. Established morphology, upregulated transcript levels of known early eye-field transcription factors and protein expression of standard retinal progenitor markers confirmed the optic vesicle/presumptive optic cup identity of <em>in vitro </em>models between day 20 and 50 of culture. This adapted protocol is relevant to researchers seeking a physiologically relevant model of early human ocular development and disease with a view to replacing animal models.</p>
Fourier-transform Infrared (FT-IR) spectroscopy fingerprints subpopulations of extracellular vesicles of different sizes and cellular origin
<p>Atomic Force Microscopy images of Large (LEV), Medium (MEV) and Small (SEV) Extrzcellular vesicles (EVs) from murine cell line B16 (B16-F10, ATCC CRL-647; Mus musculus, mouse; tissue: melanoma skin). Image size 8.3 x 8.3 um. Analysis mode: Tapping mode in air as described in Paolini et al. https://doi.org/10.1080/20013078.2020.1741174</p>
Low dose rate radiation induced secretion of TGF-β3 together with an activator in small extracellular vesicles modifies low dose hyper-radiosensitivity through ALK1 binding
<p>This is a collection of results from all clonogenic assays on T-47D cells used in the manuscript "Low dose rate radiation induced secretion of TGF-β3 together with an activator in small extracellular vesicles modifies low dose hyper-radiosensitivity through ALK1 binding". </p> <p>T-47D cells were subjected to various pretreatments: low dose rate priming (0.1-0.3 Gy/h for 1 hour), small extracellular vesicles from irradiated or control cells, irradiated or control cell conditioned medium, MMP/ADAM inhibitor TAPI-2, recombinant TGF-B3, inhibitors of ALK1, ALK2, ALK5 or TGF-BRII, iNOS inhibitor 1400W, recombinant FKBP4, recombinant MMP14 and combinations of these. All pretreatments except low dose rate priming was administered for 24 hours. </p> <p>After pretreatments, cells were seeded to colonies and irradiated with 220 kV x-rays at a dose rate of 22.5 Gy/h or gamma rays from a Co-60 source at a dose rate of 20-25 Gy/h. </p> <p>Colonies were cultured for 2-3 weeks before fixation and manual counting. </p>
Dataset to the publication "A unique signal sequence of the chemokine receptor CCR7 promotes package into COPII vesicles for efficient receptor trafficking"
<p>This repository accompanies the paper:</p> <p>"A unique signal sequence of the chemokine receptor CCR7 promotes package into COPII vesicles for efficient receptor trafficking".</p> <p>Organized into 14 folders it provides the data allowing the replication of all analyses and the manuscript's figures. For a more convenient download files were zipped.</p> <p>Upon publication of results using this dataset, please cite the following paper:</p> <p>Uetz-von Allmen E, Rippl AV, Farhan H, Legler DF. 2018. A unique signal sequence of the chemokine receptor CCR7 promotes package into COPII vesicles for efficient receptor trafficking. J Leukoc Biol 104(2):375-389.</p>
DATA: Diversity of extracellular vesicles derived from calli, cell culture and apoplastic fluid of tobacco
<p><span>The data includes .jpg files with fluorescence readings and .csv files with results of concentration and size measurements.</span></p>
Data of hybrid vesicles fusion for Nano Letters' journal article
<p>Dataset to accompany the manuscript "Thermoplasmonic induced vesicle fusion for investigating membrane protein phase affinity"</p>
Quantification of Giant Unilamellar Vesicle Fusion Products by High-Throughput Image Analysis - Microscopy Data
<p>Microscopy dataset of multipoint-multichannel images of giant unilamellar vesicles (GUVs) suspensions analysed in "Quantification of Giant Unilamellar Vesicle Fusion Products by High-Throughput Image Analysis" (under revision).</p> <p>Three folders concerning different sections of the work are included. "preliminary analysis.zip" contians the raw files and analysis scripts for recall computation and imaging setup optimization as described in the paper. Timelapse data was excluded due to file size restrictions (available upon request at the corresponding authors of the work). "IFC comparison.zip" contains raw files and analysis scripts used to optimize colocalization computation in lipid exchange and content exchange experiments. "GUV fusion analysis" contains raw files and analysis scripts for the quantification of lipid and content exchange upon sodium chloride-induced aggregation.</p> <p>Further details on the analysis are provided in the paper. The R scripts require files saved upon analysis of the raw files by the ImageJ macro "CE_analysis_CPU.ijm" included here. The R environment of the complete analysis are included in each folder to provide easier access to the elaborated data.</p>
Supplementary data for calcium-vesicles perform active diffusion in the sea urchin embryo during larval biomineralization
<p><strong>Supplementary datasets for the paper Calcium-vesicles perform active diffusion in the sea urchin embryo during larval biomineralization.</strong></p> <p>Two deskewed and deconvolved lattice light-sheet datasets (100 frames each) from the live-cell experiments are available, a control embryo dataset (01-07-2016_TimeLapse4_DMSO_21hrs_Calcein_FM464) and a VEGFR inhibited dataset (24-06-2016_Timelapse1_Axtinib_150_19hrs_Calcein_FM464). These datasets were used for collecting size and motion statistics. The control embryo dataset is available in raw microscope output without deskew or deconvolution applied (Raw_01-07-2016_TimeLapse4_DMSO_21hrs_Calcein_FM464).</p> <p>Four confocal datasets from the cytoskeletal remodeling experiments are also included, phalloidin stained images, control (Phalloidin PMC DMSO 5 zoom4s) and VEGFR inhibited (Phalloidin PMC Axt 18 zoom4); and myosinIIP stained images, control (Myosin PMC 30h DMSO new slid 4a zoom4) and VEGFR inhibited (Phalloidin PMC Axt 18 zoom4).</p> <p>Source code and instructions for the analysis tools used for both the lattice light-sheet and confocal data is available at: <a href="https://git-bioimage.coe.drexel.edu/opensource/llsm-calcium-vesicles-lever">https://git-bioimage.coe.drexel.edu/opensource/llsm-calcium-vesicles-lever</a></p> <p>Code for the deconvolution and deskew algorithms is available from the Janelia research center at: <a href="https://www.janelia.org/open-science/lattice-light-deconvolution-software-cudadeconv">https://www.janelia.org/open-science/lattice-light-deconvolution-software-cudadeconv</a></p> <p> </p> <p> </p>
Unique dynamics and exocytosis properties of GABAergic synaptic vesicles revealed by three-dimensional single vesicle tracking
<p>This data set includes x, y, and z trajectories of all GABAergic synaptic vesicles that we used for the study. These GABAergic synaptic vesicles in inhibitory presynaptic terminals of living primary hippocampal neurons were labeled by single quantum dots (QDs) conjugated with anti-VGAT antibody under electrical stimulation, and were tracked three-dimensionally by using a dual-focus imaging in real-time. Each trajectory data indicates x, y, and z positions (nanometer-scale) over time from the start of imaging to the moment of vesicle fusion. The electrical stimulation to the neurons was applied for 120 s, starting from 20 s.</p>
Raw data for "Host-interactor screens of Phytophthora infestans RXLR proteins reveal vesicle trafficking as a major effector-targeted process"
<p>This dataset contains raw and original images, phylogenetic tree files, sequence alignment files used for phylogenetic tree construction and unprocessed data for figures presented in the manuscript titled "Host-interactor screens of <em>Phytophthora infestans</em> RXLR proteins reveal vesicle trafficking as a major effector-targeted process". Each zip file contains raw data for each figure in the manuscript. A version of the manuscript is available on bioRxiv with doi.org/10.1101/2020.09.24.308585.</p>
Dataset for article: Antimicrobial peptide induced colloidal transformations in bacteria-mimetic vesicles: Combining in silico tools and experimental methods
<p><strong>Dataset for publication:</strong></p><p>Antimicrobial peptide induced colloidal transformations in bacteria-mimetic vesicles: Combining in silico tools and experimental methods<br><i>Rafael V.M. Freire, Yeny Pillco-Valencia, Gabriel C.A. da Hora, Madeleine Ramstedt, Linda Sandblad, Thereza A. Soares, Stefan Salentinig</i><br>Journal of Colloid and Interface Science Volume 596, 15 August 2021, Pages 352-363 https://doi.org/10.1016/j.jcis.2021.03.060</p><p>Setup and conditions for the experiments are described in the experimental section of the published (open access) manuscript.</p><p>Data description in README.txt file.</p>
Extracellular Vesicles Analysis in the COVID-19 Era: Insights on Serum Inactivation Protocols towards Downstream Isolation and Analysis
<p>Representative AFM images of the samples analyzed in the relative manuscript. Raw data just imported from the AFM multimode native format to the Gwyddion Open source data analysis software</p>
Small extracellular vesicles from malignant ascites of patients with advanced ovarian cancer provide insights into the dynamics of the extracellular matrix
<p>Representative Atomic Force Microscope images collected in liquid on Extracellular Vesicles from Ascite and Cell cultures. Data were collected by Broker Multimode VIII. Images like these were used in the paper to measure the contact angle and size distribution.</p>
Ultrafst whole cell imaging data of SiT-Golgi vesicles over 10,000 time points
<p>This dataset is the raw lattice light-sheet microscopy data of SiT-Golgi vesicles over 10,000 consecutive time points, which was used to demonstrate TiS-rDL denoising algorithm in our Nature Biotechnology paper (DOI: 10.1038/s41587-022-01471-3). This dataset can be used for non-commercial purposes with proper citations of our NBT paper.</p>
Vesicle size analysis dataset of Eldgjá tephra
<p>Raw vesicle area and metadata acquired by image analysis of thin-sections of Eldgjá tephra clasts. Acquired using the methods outlined by <a href="https://doi.org/10.1016/j.jvolgeores.2009.12.003">Shea et al. (2010)</a>, <a href="https://doi.org/10.1016/S0377-0273(98)00043-2">Sahagian & Proussevitch (1998)</a>, and <a href="https://hdl.handle.net/20.500.11815/324">Moreland (2019a)</a>.</p> <p>The format is compatible with the VSA Processor script (<a href="https://doi.org/10.5281/zenodo.2591179">Moreland 2019b</a>).</p>
Dataset related to article "Glia-to-neuron transfer of miRNAs via extracellular vesicles: a new mechanism underlying inflammation-induced synaptic alterations"
<p>This record contains raw data related to article "Glia-to-neuron transfer of miRNAs via extracellular vesicles: a new mechanism underlying inflammation-induced synaptic alterations"</p> <p>Recent evidence indicates synaptic dysfunction as an early mechanism affected in neuroinflammatory diseases, such as multiple sclerosis, which are characterized by chronic microglia activation. However, the mode(s) of action of reactive microglia in causing synaptic defects are not fully understood. In this study, we show that inflammatory microglia produce extracellular vesicles (EVs) which are enriched in a set of miRNAs that regulate the expression of key synaptic proteins. Among them, miR-146a-5p, a microglia-specific miRNA not present in hippocampal neurons, controls the expression of presynaptic synaptotagmin1 (Syt1) and postsynaptic neuroligin1 (Nlg1), an adhesion protein which play a crucial role in dendritic spine formation and synaptic stability. Using a Renilla-based sensor, we provide formal proof that inflammatory EVs transfer their miR-146a-5p cargo to neuron. By western blot and immunofluorescence analysis we show that vesicular miR-146a-5p suppresses Syt1 and Nlg1 expression in receiving neurons. Microglia-to-neuron miR-146a-5p transfer and Syt1 and Nlg1 downregulation do not occur when EV-neuron contact is inhibited by cloaking vesicular phosphatidylserine residues and when neurons are exposed to EVs either depleted of miR-146a-5p, produced by pro-regenerative microglia, or storing inactive miR-146a-5p, produced by cells transfected with an anti-miR-146a-5p. Morphological analysis reveals that prolonged exposure to inflammatory EVs leads to significant decrease in dendritic spine density in hippocampal neurons in vivo and in primary culture, which is rescued in vitro by transfection of a miR-insensitive Nlg1 form. Dendritic spine loss is accompanied by a decrease in the density and strength of excitatory synapses, as indicated by reduced mEPSC frequency and amplitude. These findings link inflammatory microglia and enhanced EV production to loss of excitatory synapses, uncovering a previously unrecognized role for microglia-enriched miRNAs, released in association to EVs, in silencing of key synaptic genes.</p>
Dataset related to article "Decoding distinctive features of plasma extracellular vesicles in amyotrophic lateral sclerosis"
<p>The mass spectrometry proteomics data have been deposited at the ProteomeXchange Consortium (<a href="http://proteomecentral.proteomexchange.org/cgi/GetDataset">http://proteomecentral.proteomexchange.org/cgi/GetDataset</a>) via the PRIDE partner repository with the data set identifier PXD020629.</p>
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Allen Brain Atlas
Allen Brain Atlas is an Allen Institute collection of brain map atlases, datasets, APIs, and analysis tools covering mouse, human, and non-human primate brain resources.
Annotated Behaviour and Observability Dataset (ABODe)
ABODe is a University of Edinburgh DataShare dataset for behavior classification in group-housed mice using home-cage video, identities, bounding boxes, ground-plate positions, and annotator labels.
DANDI Archive for NWB datasets
DANDI is a BRAIN Initiative archive for publishing and sharing neurophysiology data, including electrophysiology, optophysiology, and behavioral data packaged as NWB and related standards.
International Brain Laboratory public data
The International Brain Laboratory public data releases expose standardized mouse decision-making experiments, including Neuropixels recordings, widefield calcium imaging, behavior, and session metadata accessed through the ONE API.
OpenNeuro
OpenNeuro is a free, open platform for sharing neuroimaging datasets, with public search, dataset pages, and download paths for web, S3, DataLad, and the OpenNeuro CLI.