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Example Microscopy Metadata JSON files produced using Micro-Meta App to document example microscopy experiments performed at individual core facilities
<p>Example <strong>Microscopy Metadata </strong>(Microscope.JSON and Settings.JSON)<strong> files </strong>produced using<strong> <a href="https://wu-bimac.github.io/MicroMetaApp.github.io/">Micro-Meta App</a> </strong>to document the <strong>Hardware Specifications</strong> of example Microscopes and the <strong>Image Acquisition Settings</strong> utilized to acquire example images as listed in the table below.</p> <blockquote> <p>For each facility, the dataset contains two JSON files:</p> <ol> <li><strong>Microscope.JSON file</strong> (e.g., 01_marcello_uliverpool_cci_zeiss_axioobserz1_lsm710.json)</li> <li><strong>Settings.JSON file</strong> (indicated with the name of the image and with the _AS suffix)</li> </ol> </blockquote> <p><strong>Micro-Meta App was</strong> developed as part of a <strong>global community initiative</strong> including the <a href="http://www.4dnucleome.org/"><strong>4D Nucleome (4DN)</strong> </a>Imaging Working Group, <strong>BioImaging North America (BINA)</strong> <a href="https://www.bioimagingna.org/qc-dm-wg">Quality Control and Data Management Working Group</a>, and <strong>QUAlity and REProducibility for Instrument and Images in Light Microscopy</strong> (<a href="https://quarep.org/"><strong>QUAREP-LiMi</strong></a>), to extend the <strong>Open Microscopy Environment (OME)</strong> <a href="https://www.openmicroscopy.org/Schemas/Documentation/Generated/OME-2016-06/ome.html">data model</a>.</p> <blockquote> <p>The works of this <strong>global community effort</strong> resulted in multiple publications featured on a recent <strong>Nature Methods FOCUS ISSUE </strong>dedicated to <a href="https://www.nature.com/collections/djiciihhjh">Reporting and reproducibility in microscopy</a>.</p> </blockquote> <blockquote> <p><strong>Learn More!</strong> For a thorough description of <strong>Micro-Meta App</strong> consult our recent <a href="https://doi.org/10.1038/s41592-021-01315-z">Nature Methods</a> and <a href="https://doi.org/10.1101/2021.05.31.446382">BioRxiv.org</a> publications!</p> </blockquote> <p> </p> <table> <tbody> <tr> <td><strong>Nr.</strong></td> <td><strong>Manufacturer</strong></td> <td><strong>Model</strong></td> <td><strong>Tier</strong></td> <td><strong>Εxperiment Type</strong></td> <td><strong>Facility Name</strong></td> <td><strong>Department and Institution</strong></td> <td><strong>URL</strong></td> <td><strong>References</strong></td> </tr> <tr> <td>1</td> <td><strong>Carl Zeiss Microscopy</strong></td> <td><strong>Axio Observer Z1 (with LSM 710 scan head)</strong></td> <td>1</td> <td>3D visualization of superhydrophobic polymer-nanoparticles</td> <td>Centre for Cell Imaging (CCI)</td> <td>University of Liverpool</td> <td>https://cci.liv.ac.uk/equipment_710.html</td> <td>Upton et al., 2020</td> </tr> <tr> <td>2</td> <td><strong>Carl Zeiss Microscopy</strong></td> <td><strong>Axio Observer (Axiovert 200M)</strong></td> <td>2</td> <td>Μeasurement of illumination stability on Chinese Hamster Ovary cells expressing Paxillin-EGFP</td> <td>Advanced BioImaging Facility (ABIF).</td> <td>McGill University</td> <td>https://www.mcgill.ca/abif/equipment/axiovert-1</td> <td>Kiepas et al., 2020</td> </tr> <tr> <td>3</td> <td><strong>Carl Zeiss Microscopy</strong></td> <td><strong>Axio Observer Z1 (with Spinning Disk)</strong></td> <td>2</td> <td>Immunofluorescence imaging of cryosection of Mouse kidney</td> <td>Imagerie Cellulaire; Quality Control managed by Miacellavie (https://miacellavie.com/)</td> <td>Centre de recherche du Centre Hospitalier Université de Montréal (CR CHUM), University of Montreal</td> <td>https://www.chumontreal.qc.ca/crchum/plateformes-et-services (the web site is for all core facilities, not specifically for the core facility hosting this microscope)</td> <td>Pilliod et al., 2020</td> </tr> <tr> <td>4</td> <td><strong>Carl Zeiss Microscopy</strong></td> <td><strong>Axio Imager Z2 (with Apotome)</strong></td> <td>2</td> <td>Immunofluorescence imaging of mitotic division in Hela cells using </td> <td>Bioimaging Unit</td> <td>Newcastle University</td> <td>https://www.ncl.ac.uk/bioimaging/</td> <td>Watson et al., 2020</td> </tr> <tr> <td>5</td> <td><strong>Carl Zeiss Microscopy</strong></td> <td><strong>Axio Observer Z1</strong></td> <td>2</td> <td>Fluorescence microscopy of human skin fibroblasts from Glycogen Storage Disease patients.</td> <td>Life Imaging Center (LIC)</td> <td>Centre for Integrative Signalling Analysis (CISA), University of Freiburg</td> <td>https://miap.eu/equipments/sd-i-abl/</td> <td>Hannibal et al., 2020</td> </tr> <tr> <td>6</td> <td><strong>Leica Microsystems</strong></td> <td><strong>DMI6000B</strong></td> <td>2</td> <td>3D immunofluorescence imaging rhinovirus infected macrophages </td> <td>IMAG'IC Confocal Microscopy Facility</td> <td>Institut Cochin, CNRS, INSERM, Université de Paris</td> <td>https://www.institutcochin.fr/core_facilities/confocal-microscopy/cochin-imaging-photonic-microscopy/organigram_team/10054/view</td> <td>Jubrail et al., 2020</td> </tr> <tr> <td>7</td> <td><strong>Leica Microsystems</strong></td> <td><strong>DM5500B</strong></td> <td>2</td> <td>Immunofluorescence analysis of the colocalization of PML bodies with DNA double-strand breaks</td> <td>Bioimaging Unit</td> <td>Edwardson Building on the Campus for Ageing and Vitality, Newcastle University</td> <td>https://www.ncl.ac.uk/bioimaging/equipment/leica-dm5500/#overview</td> <td>da Silva et al., 2019; Nelson et al., 2012<br> </td> </tr> <tr> <td>8</td> <td><strong>Leica Microsystems</strong></td> <td><strong>DMI8-CS (with TCS SP8 STED 3X)</strong></td> <td>2</td> <td>Live-cell imaging of N. benthamiana leaves cells-derived protoplasts</td> <td>Center for Advanced Imaging (CAi)</td> <td>School of Mathematics/Natural Sciences, Heinrich-Heine-Universität Düsseldorf</td> <td>https://www.cai.hhu.de/en/equipment/super-resolution-microscopy/leica-tcs-sp8-sted-3x</td> <td>Singer et al., 2017; Hänsch et al., 2020</td> </tr> <tr> <td>9</td> <td><strong>Nikon Instruments</strong></td> <td><strong>Eclipse Ti</strong></td> <td>2</td> <td>Immunofluorescence analysis of the cytoskeleton structure in COS cells</td> <td>Advanced Imaging Center (AIC)</td> <td>Janelia Research Campus, Howard Hughes Medical Institute</td> <td>https://www.janelia.org/support-team/light-microscopy/equipment</td> <td>Abdelfattah et al., 2019; Qian et al., 2019; Grimm et al., 2020</td> </tr> <tr> <td>10</td> <td><strong>Nikon Instruments</strong></td> <td><strong>Eclipse Ti-E (HCA)</strong></td> <td>2</td> <td>Τime-lapse analysis of the bursting behavior of amine-functionalized vesicular assemblies</td> <td>Light Microscopy Facility (IALS-LIF)</td> <td>Institute for Applied Life Sciences, University of Massachusetts at Amherst</td> <td>https://www.umass.edu/ials/light-microscopy</td> <td>Fernandez et al., 2020</td> </tr> <tr> <td>11</td> <td><strong>Nikon Instruments/Coleman laboratory (customized)</strong></td> <td><strong>TIRF HILO Epifluorescence light Microscope (THEM)/ Eclipse Ti</strong></td> <td>2</td> <td>Single-particle tracking of Halo-tagged PCNA in Lox cells</td> <td>Coleman laboratory</td> <td>Anatomy and Structural Biology Department, The Albert Einstein College of Medicine</td> <td>https://einsteinmed.org/faculty/12252/robert-coleman/</td> <td>Drosopoulos et al., 2020</td> </tr> <tr> <td>12</td> <td><strong>Nikon Instruments</strong></td> <td><strong>Eclipse Ti (with Andor Dragon Fly Spinning Disk)</strong></td> <td>2</td> <td>Investigation of the 3D structure of cerebral organoids</td> <td>Montpellier Resources Imagerie</td> <td>Centre de Recherche de Biologie cellulaire de Montpellier (MRI-CRBM), CNRS, Univerity of Montpellier</td> <td>https://www.mri.cnrs.fr/en/optical-imaging/our-facilities/mri-crbm.html</td> <td>Ayala-Nunez et al., 2019</td> </tr> <tr> <td>13</td> <td><strong>Nikon Instruments</strong></td> <td><strong>Eclipse Ti2</strong></td> <td>2</td> <td>Ιmmunofluorescence imaging of cryosections of mouse hearth myocardium </td> <td>Neuroscience Center Microscopy Core</td> <td>Neuroscience Center, University of North Carolina</td> <td>https://www.med.unc.edu/neuroscience/core-facilities/neuro-microscopy/</td> <td>Aghajanian et al., 2021</td> </tr> <tr> <td>14</td> <td><strong>Nikon Instruments</strong></td> <td><strong>Eclipse Ti2</strong></td> <td>2</td> <td>Live-cell imaging of bacterial cells expressing GFP-PopZ</td> <td>Microscopy Resources on the North Quad (MicRoN)</td> <td>Harvard Medical School </td> <td>https://micron.hms.harvard.edu/</td> <td>Lim and Bernhardt 2019; Lim et al., 2019</td> </tr> <tr> <td>15</td> <td><strong>Olympus/Biomedical Imaging Group (customized)</strong></td> <td><strong>TIRF Epifluorescence Structured light Microscope (TESM)/IX71</strong></td> <td>3</td> <td>3D distribution of HIV-1 in the nucleus of human cells</td> <td>Biomedical Imaging Group</td> <td>Program in Molecular Medicine, University of Massachusetts Medical School</td> <td>https://trello.com/b/BQ8zCcQC/tirf-epi-fluorescence-structured-light-microscope</td> <td>Navaroli et al., 2012</td> </tr> <tr> <td>16</td> <td><strong>Olympus/Computer Vision Laboratory (customized)</strong></td> <td><strong>3D BrightField Scanner/IX71</strong></td> <td>3</td> <td>Transmitted light brightfield visualization of swimming spermatocytes</td> <td>Laboratorio Nacional de Microscopia Avanzada (LNMA) and Computer Vision Laboratory of the Institute of Biotechnology</td> <td>Universidad Nacional Autonoma de Mexico (UNAM)</td> <td>https://lnma.unam.mx/wp/</td> <td>Pimentel et al., 2012; Silva-Villalobos et al., 2014</td> </tr> </tbody> </table> <p><strong>Getting started</strong></p> <p>Use these videos to get started with using Micro-Meta App after installation into OMERO and downloading the example data files:</p> <ol> <li><a href="https://vimeo.com/562022222">Video 1</a></li> <li><a href="https://vimeo.com/562022281">Video 2</a></li> </ol> <p><strong>More information</strong></p> <blockquote> <p>For full information on how to use Micro-Meta App please utilize the following resources:</p> <ol> <li>Micro-Meta App <a href="https://wu-bimac.github.io/MicroMetaApp.github.io/">website</a></li> <li><a href="https://micrometaapp-docs.readthedocs.io/en/latest/index.html">Full documentation</a></li> <li><a href="https://micrometaapp-docs.readthedocs.io/en/latest/docs/intro/installation.html">Installation</a> instructions</li> <li><a href="https://micrometaapp-docs.readthedocs.io/en/latest/docs/tutorials/index.html#step-by-step-instructions">Step-by-Step Instructions</a></li> <li><a href="https://micrometaapp-docs.readthedocs.io/en/latest/docs/tutorials/VideoTutorials.html#micro-meta-app-video-tutorials">Tutorial Videos</a></li> </ol> </blockquote> <p><strong>Background</strong></p> <p>If you want to learn more about the importance of <strong>metadata and quality contro</strong>l to ensure full <strong>reproducibility, quality and scientific value</strong> in light microscopy, please take a look at our recent publications describing the development of community-driven light <strong>4DN-BINA-OME Microscopy Metadata</strong> specifications <a href="https://doi.org/10.1038/s41592-021-01327-9">Nature Methods</a> and <a href="https://doi.org/10.1101/2021.04.25.441198">BioRxiv.org</a> and our <a href="https://arxiv.org/abs/1910.11370">overview manuscript</a> entitled <strong>A perspective on Microscopy Metadata: data provenance and quality control</strong>.</p> <p> </p> <p> </p>
Example Microscopy Metadata JSON files produced using Micro-Meta App to document the acquisition of example images using a custom-built TIRF Epifluorescence Structured Illumination Microscope
<p><strong>Example Microscopy Metadata JSON files produced using the <a href="https://wu-bimac.github.io/MicroMetaApp.github.io/">Micro-Meta App</a> documenting an example raw-image file acquired using the custom-built TIRF Epifluorescence Structured Illumination Microscope.</strong></p> <p>For this use case, which is presented in Figure 5 of <a href="http://doi: https://doi.org/10.1101/2021.05.31.446382">Rigano et al., 2021</a>, Micro-Meta App was utilized to document:</p> <p>1) The <strong>Hardware Specifications</strong> of the custom build TIRF Epifluorescence Structured light Microscope (TESM; <a href="https://www.pnas.org/content/109/8/E471.long">Navaroli et al., 2010</a>) developed, built on the basis of the based on Olympus IX71 microscope stand, and owned by the Biomedical Imaging Group (http://big.umassmed.edu/) at the Program in Molecular Medicine of the University of Massachusetts Medical School. Because TESM was custom-built the most appropriate documentation level is <strong>Tier 3</strong> (<em>Manufacturing/Technical Development/Full Documentation</em>) as specified by the <a href="https://doi.org/10.5281/zenodo.4710731">4DN-BINA-OME</a> Microscopy Metadata model (<a href="https://doi.org/10.1101/2021.04.25.441198">Hammer et al., 2021</a>).</p> <p>The TESM Hardware Specifications are stored in: <strong>Rigano et al._Figure 5_UseCase_Biomedical Imaging Group_TESM.JSON</strong></p> <p>2) The <strong>Image Acquisition Settings</strong> that were applied to the TESM microscope for the acquisition of an example image (FSWT-6hVirus-10minFIX-stk_4-EPI.tif.ome.tif) obtained by Nicholas Vecchietti and Caterina Strambio-De-Castillia. For this image, TZM-bl human cells were infected with HIV-1 retroviral three-part vector (FSWT+PAX2+pMD2.G). Six hours post-infection cells were fixed for 10 min with 1% formaldehyde in PBS, and permeabilized. Cells were stained with mouse anti-p24 primary antibody followed by DyLight488-anti-Mouse secondary antibody, to detect HIV-1 viral Capsid. In addition, cells were counterstained using rabbit anti-Lamin B1 primary antibody followed by DyLight649-anti-Rabbit secondary antibody, to visualize the nuclear envelope and with DAPI to visualize the nuclear chromosomal DNA.</p> <p>The Image Acquisition Settings used to acquire the FSWT-6hVirus-10minFIX-stk_4-EPI.tif.ome.tif image are stored in: <strong>Rigano et al._Figure 5_UseCase_AS_fswt-6hvirus-10minfix-stk_4-epi.tif.JSON</strong></p> <p><em><strong>Instructional video tutorials on how to use these example data files:</strong></em><br> Use these videos to get started with using Micro-Meta App after downloading the example data files available here.</p> <ul> <li><a href="https://vimeo.com/562022222">Part 1/2</a></li> <li><a href="https://vimeo.com/562022281">Part 2/2</a></li> </ul>
Interlaboratory study: Testing reproducibility of solid biofuels component identification using reflected light microscopy
<p><strong>Submitted data was used to write an article: </strong>Drobniak, A., Mastalerz, M., Jelonek, Z., Jelonek, I., Adsul, T., Andolšek, N., Ardakani, O.H., Congo, T., Demberelsuren, B., Donohoe, B.S., Douds, A., Flores, D., Ganzorig, R., Ghosh, S., Gize, A., Goncalves, P.A., Hackely, P., Hatcherian, J., Hower, J.C., Kalaitzidis, S., Kędzior, S., Knowles, W., Kuś, J., Lis, K., Lis, G., Liu, B., Luo, Q., Du, M., Mishra, D., Misz-Kennan, M., Mugerwa, T., O'Keefe, J., Park, J., Pearson, R., Petersen, H., Reyes, J., Ribeiro, J., Niedzwiedzkas, J.L., de la Rosa Rodriguez, G., Sosnowski, P., Valentine, B., Varma, A., Wojtaszek-Kalaitzidi, M., Xu, Z., Zdravkov, A., Ziemianin, K., Interlaboratory study: Testing reproducibility of biomass fuels component identification using reflected light microscopy. International Journal of Coal Geology 277, 104331. <a href="https://doi.org/10.1016/j.coal.2023.104331">https://doi.org/10.1016/j.coal.2023.104331</a>.</p> <p> </p> <p><strong>Funding acknowledgments: </strong>The project is co-financed by the Polish National Agency for Academic Exchange within the Polish Returns Programme (BPN/PPO/2021/1/00005/DEC/1), the National Science Center, Poland (2022/01/1/ST10/00024), and the research activities co-financed by the funds granted under the Research Excellence Initiative of the University of Silesia in Katowice, Poland. </p> <p> </p> <p><strong>Article Abstract: </strong>Considering global market trends and concerns about climate change and sustainability, increased biomass use for energy is expected to continue. As more diverse materials are being utilized to manufacture solid biomass fuels, it is critical to implement quality assessment methods to analyze these fuels thoroughly. One such method is reflected light microscopy (RLM), which has the potential to complement and enhance current standard testing, leading to improving fuel quality assessment and, ultimately, preventing avoidable air pollution. An interlaboratory study (ILS) was conducted to test the reproducibility of biomass fuels component identification using a reflected light microscopy technique. The exercise was conducted on thirty photomicrographs showing biomass and various undesired components (like plastics or mineral matter), which were purposely added (by the ILS organizers) to contaminate wood pellets and charcoal-based grilling fuels. Forty-six participants had various levels of difficulty identifying the marked components, and as a result, the percentage of correct answers ranged from 52.2 to 94.4%. Among the most difficult components to distinguish were petroleum products and inorganic matter. Various reasons led to the misidentification, including insufficient morphological descriptions of the components provided to participants, ambiguities of the nomenclature, limitations of the analytical and exercise method, and insufficient experience of the participants. Overall, the results indicate that RLM has the potential to enhance the quality assessment of biomass fuels. However, they also demonstrate that the petrographic classification used in this exercise requires further refinement before it can be standardized. While a new simplified classification of solid biomass fuels components was created as an outcome of this study, future research is necessary to refine the nomenclature, develop a microscopic morphological description of the components, and verify the accuracy of component identification with a follow-up ILS.</p>
Raw Data on Extracellular Particles in 613 Human and 163 Canine Diluted Plasma and Blood Samples Assessed by Interferometric Light Microscopy
<p><span>Extracellular nanoparticles (EPs) are cellular fragments. After being released in cell exterior, they become mediators of the cell-cell interaction. Their characterization in bodily fluids may reflect the clinical status of the organism. Here we present data on the number density <em>n</em> and hydrodynamic diameter <em>D</em><sub>h </sub>of EPs assessed directly in diluted plasma and blood by using a recently developed technique, Interferometric Light Microscopy (Romolo et al., 2022). The data are presented in the attached Table. </span></p> <p><span>We collected 613 blood and plasma samples from human patients with Inflammatory Bowel Disease (IBD) taken into tubes with trisodium citrate and ethylenediaminetetraacetic acid (EDTA) anticoagulants and 163 blood and plasma samples from canine patients with Brachycephalic Obstructive Airway Syndrome (BOAS). </span><span>The human study was conducted in accordance with the Declaration of Helsinki, and approved by the National Medical Ethics Committee of the Republic of Slovenia (0120-271/2022/4; KME 27 July 2022). All procedures in the animal study complied with the relevant Slovenian government regulations (Animal Protection Act, Official Gazette of the Republic of Slovenia, No. 43/2007). The animal study was approved by the Animals in Experiments Welfare Commission of the Veterinary Faculty, University of Ljubljana, approval number 18-3/2022-1. </span><span>Information regarding sample preparation is documented in the MIBlood-EV reports.</span></p> <div> <div> <div><span><a name="_msocom_1"></a></span></div> </div> </div>
Atomic force microscopy indentation data of zebrafish spinal cord sections
<p>The HDF5 file was created using the Python package nanite. It contains 1132 raw atomic force microscopy (AFM) force-indentation curves of zebrafish spinal cord sections, the preprocessed curves, and the corresponding fits to the approach part. In addition, a manual rating was assigned to each force-indentation curve. The intended use of this dataset is the application of machine-learning approaches to quantify AFM data quality for biological tissues.</p>
In vivo rat brain for Ultrasound Localization Microscopy: raw and beamformed data.
<p><strong>Datasets provided for Open Platform for Ultrasound Localization Microscopy: Performance Assessment of Localization Algorithms.</strong></p> <p><strong>Abstract:</strong></p> <p>Ultrasound Localization Microscopy (<strong>ULM</strong>) is an ultrasound imaging technique that relies on the acoustic response of sub-wavelength ultrasound scatterers to map the microcirculation with an order of magnitude increase in resolution. Initially demonstrated <em>in vitro</em>, this technique has matured and sees implementation<em> in vivo</em> for vascular imaging of organs, and tumors in both animal models and humans. The performance of the localization algorithm greatly defines the quality of vascular mapping. We compiled and implemented a collection of ultrasound localization algorithms and devised three datasets<em> in silico</em> and<em> in vivo</em> to compare their performance through 18 metrics. We also present two novel algorithms designed to increase speed and performance. By openly providing a complete package to perform ULM with the algorithms, the datasets used, and the metrics, we aim to give researchers a tool to identify the optimal localization algorithm for their usage, benchmark their software and enhance the overall image quality in the field while uncovering its limits.</p> <p>This article provides all materials and post-processing scripts and functions.</p> <p><strong>Methods:</strong></p> <p>200.000 ultrasound images have been acquired <em>in vivo </em>on a rat brain with skull removal at 1000 Hz with a 15 MHz linear probe.</p> <p>This dataset contains raw radiofrequency data (<strong>RF</strong>) and beamformed images (<strong>IQ</strong>) of the brain vascularization with flowing microbubbles (ultrasound contrast agent).</p> <p><strong>Article to be cited:</strong> Heiles, Chavignon, Hingot, Lopez, Teston and Couture.<br> <a href="http://doi.org/10.1038/s41551-021-00824-8"><em>Performance benchmarking of microbubble-localization algorithms for ultrasound localization microscopy</em>, Nature Biomedical Engineering, 2022, (doi.org/10.1038/s41551-021-00824-8)</a>.</p> <p><strong>Related processing scripts and codes:</strong> <a href="https://github.com/AChavignon/PALA">github.com/AChavignon/PALA</a></p> <p><strong>Related datasets:</strong> <a href="https://doi.org/10.5281/zenodo.4343435">doi.org/10.5281/zenodo.4343435</a></p> <p><strong>Acknowledgments:</strong></p> <p>We thank Cyrille Orset (INSERM UMR-S U1237, Physiopathology and Imaging of Neurological Disorders, GIP Cyceron, BB@C, Caen, France) for animals’ preparation and perfusion of contrast agent and the biomedical imaging platform CYCERON (UMS 3408 Unicaen/CNRS, Caen, France).</p>
Synthetic images of cell nuclei in widefield microscopy
<p>The images were generated by <a href="http://www.cs.tut.fi/sgn/csb/simcep/tool.html">SIMCEP</a>, a widefield fluorescence microscopy biological images simulator.</p> <p>The dataset is used to demonstrate the execution of image analysis workflows with BIAFLOWS on a local machine from a jupyter notebook.</p>
Example dataset for single-photon microscopy
<p>Datasets for spot-variation fluorescence fluctuation analysis, fluorescence lifetime fluctuation analysis and fluorescence cross-correlation analysis. One dataset is intensity-based, while three are time-tagged based.</p>
Supplementary Materials for "Accelerating data sharing and re-use in volume electron microscopy"
<p>The deposition contains supporting materials for "Accelerating data sharing and re-use in volume electron microscopy" Comment</p> <ul> <li>Sample preparation protocol for cell monolayers optimized for serial block face scanning electron microscopy</li> <li>Supporting movies showing models of biological specimens imaged using volume electron microscopy</li> </ul>
Raw data from the manuscript "Full-aperture extended-depth oblique plane microscopy through dynamic remote focusing"
<p>The repository contains all the raw data from the manuscript titled "Full-aperture extended-depth oblique plane microscopy through dynamic remote focusing" (https://doi.org/10.1117/1.JBO.29.3.036502).<br> The data consists in 3D stacks acquired with the method described in the manuscript. Since raw images are acquired along a diagonal plane, and are stretched in one direction, the dataset also includes a Python script to perform an affine transform projecting the stack on cartesian coordinates.</p> <p>Samples imaged include sub-resolution microbeads in agarose gel, a fixed slice of mouse kidney (fluocells prepared slide #3, invitrogen), and 3 to 5 days post fertilization Tg(kdrl:eGFP)s843 Zebrafish.</p>
Confocal Microscopy Visualizes Particle-Crack Interactions in Epoxy Composites with Optical Force Probe-Crosslinked Rubber Particles
<p>Data (*.csv and *.lif) corresponding to Figures 2-7 of the manuscript and Figures S1-S2 of the Supporting Information.</p>
Mechanical characterisation of the developing cell wall layers of tension wood fibres by Atomic Force Microscopy
<p>This dataset corresponds to the Arnould et al. (2022) paper (available at https://www.biorxiv.org/content/10.1101/2021.09.23.461481v1.full) on the mechanical characterization of developing cell wall layers of tension wood fibers by Atomic Force Microscopy. It contains all raw AFM files (Bruker format .spm, readable by the free software Gwyddion for example) corresponding to mechanical measurements of poplar reaction wood cells (clone 717-1B4) along 3 radial lines/rows, starting from the cambium. Each cell is identified by its "macroscopic" distance from the cambium (value in µm in the name of each file corresponding to the displacement of the sample in the AFM) which was corrected after using the AFM optical image captures. Some files, with a -z extension after the distance value, correspond to a zoom into the cell wall. The data also contain measurements made for mechanical calibration on epoxy embedded Kevlar fibers, controlled measurements in the embedding resin between each radial line and measurements in normal wood cells. Two csv files containing final data extracted from AFM measurements that give the value of the indentation modulus and the relative thickness to cell diameter ratio (by AFM and by phase contrast optical microscopy) in each cell wall layer as a function of cambium distance are also provided.</p>
Bridging the gap between single nanoparticle imaging and global electrochemical response by correlative microscopy assisted by machine vision
<p>The data in this repository corresponds to experimental data: linear sweep voltammetry, optical movie and the database of the SEM images. They support the findings of a study discussed in the article by Godeffroy et al. published in Small Methods with the doi: http:/doi.org/10.1002/smtd.202200659. The data analysis to reproduce the results presented in the article has been carried out by homemade Python program routines also provided in this repository. The descirption of each routine is also provided in a text file.</p>
Mapping mineralogical heterogeneities at the nm-scale by scanning electron microscopy in modern Sardinian stromatolites: Deciphering the origin of their laminations
<p>These are the raw or processed data used for a paper published in Chemical Geology by Debrie et al. (2022), entitled "Mapping mineralogical heterogeneities at the nm-scale by scanning electron microscopy in modern Sardinian stromatolites: Deciphering the origin of their laminations", <a href="https://doi.org/10.1016/j.chemgeo.2022.121059">https://doi.org/10.1016/j.chemgeo.2022.121059</a></p> <p>The data content is summarized in the List_description_of_data.xlsx file</p>
Dataset of Scanning Tunneling Microscopy (STM) images of model surfaces for elementary steps in catalytic reactions
<p>STM images presented in the dataset were recorded by the STRAS research group using a Omicron Variable Temperature STM (VT-STM) microscope, in the TASC laboratory of the CNR-IOM in Trieste.</p> <p>This work has been done within the NFFA-DI project funded by the European Union – NextGenerationEU - Missione 4, “Istruzione e Ricerca” – Componente 2, “Dalla ricerca all'impresa” – Linea di investimento 3.1,“Fondo per la realizzazione di un sistema integrato di infrastrutture di ricerca e innovazione” – Azione 3.1.1, “Creazione di nuove IR o potenziamento di quelle esistenti che concorrono agli obiettivi di Eccellenza Scientifica di Horizon Europe e costituzione di reti”.</p>
Short-Exposure Transmission Electron Microscopy of Cilia
<p>Noisy and pseudo ground-truth short-exposure transmission electron microscopy (TEM) images of Cilia used to obtain the results depicted in Fig. 3 in the paper "Zero-Shot Denoising of Microscopy Images Recorded at High-Resolution Limits" (Salwig & Drefs et al., 2024). The images were derived based on a dataset provided upon personal communication with the authors of the paper "Denoising of Short Exposure Transmission Electron Microscopy Images For Ultrastructural Enhancement" (Bajíc et al., 2018). </p> <p>The original dataset consisted of a sequence of 100 noisy short-exposure TEM images of a scene showing a cilium. The images had a resolution of 2048 × 2048 pixels, and each image depicted a slightly shifted version of the scene. The file pseudo-ground-truth.tif was obtained by first aligning all images of the sequence using rigid registration (Marstal et al., 2016) and subsequently computing the pixel-wise median (following a procedure discussed in Bajíc et al., 2018). The file noisy.tif was obtained by randomly selecting one image from the sequence (the 91st image).</p> <p>The images are stored in 16 bit TIF format. For visualization, use an image viewer capable of reading 16 bit images (e.g. ImageJ).</p>
An annotated high-content fluorescence microscopy dataset with EGFP-Galectin-3-stained cells and manually labelled outlines
<p>Here we present a benchmarking dataset of fluorescence microscopy images with EGFP-Galectin-3-stained cells together with annotations of their outlines. Images were randomly selected from an RNA interference screen with a modified U2OS osteosarcoma cell line, acquired on a Thermo Fischer CX7 high-content imaging system at 20x magnification. </p> <p>The dataset contains 60 images showing over 2000 labelled nuclear objects in total, which is sufficiently large to train well-performing neural networks for instance or semantic segmentation. It is pre-split into training, development and test set, each in a zip file. The dataset should be referred to as Aitslab_bioimaging2.</p> <p>For most of the images, nuclear staining and annotations have been published previously in the dataset Aitslab_bioimaging1 (https://doi.org/10.5281/zenodo.6657260). The conversion script to produce the png images from the C01 images was published together with this dataset.</p>
Comparison and practical review of segmentation approaches for label-free microscopy
<p>This dataset contains microscopic images of PNT1A cell line captured by multiple microcopic without use of any labeling and a manually annotated ground truth for subsequent use in segmentation algorithms. Dataset also includes images reconstructed according to the methods described below in order to ease further segmentation. </p> <p>See Vicar et al. Cell segmentation methods for label-free contrast microscopy: review and comprehensive comparison. BMC Bioinformatics (2019) 20:360. DOI <a href="https://doi.org/10.1186/s12859-019-2880-8">10.1186/s12859-019-2880-8</a></p> <p>Code using this dataset is available at <a href="https://github.com/tomasvicar/Cell-segmentation-methods-comparison">https://github.com/tomasvicar/Cell-segmentation-methods-comparison</a></p> <p><strong>Materials and methods </strong></p> <p>Cells were cultured in RPMI-1640 medium supplemented with antibiotics (penicillin 100 U/ml and streptomycin 0.1 mg/ml) with 10% fetal bovine serum. Prior microscopy acquisition, cells were maintained at 37 cenigrade in a humidified incubator with 5% CO2. Intentionally, high passage number of cells was used (>30) in order to describe distinct morphological heterogeneity of cells (rounded and spindle-shaped, relatively small to large polyploid cells). For acquisition purposes, cells were cultivated in Flow chambers µ-Slide I Luer Family (Ibidi, Martinsried, Germany).</p> <p>Quantitative phase imaging (QPI) microscopy was performed on Tescan Q-PHASE (Tescan, Brno, Czech republic), with objective Nikon CFI Plan Fluor 10x/0.30 captured by Ximea MR4021MC (Ximea, Münster, Germany). Imaging is based on the original concept of coherence-controlled holographic microscope \cite{Kolman:10,Slaby:13}, images are shown in grayscale with units of pg/µm2.</p> <p>DIC microscopy was performed on microscope Nikon A1R (Nikon, Tokyo, Japan), with objective Nikon CFI Plan Apo VC 20x/0.75 captured by CCD camera Jenoptik ProgRes MF (Jenoptik, Jena, Germany). </p> <p>HMC microscopy was performed on microscope Olympus IX71 (Olympus, Tokyo, Japan), with objective Olympus CplanFL N 10x/0.3 RC1 captured by CCD camera Hamamatsu Photonics ORCA-R2 (Hamamatsu Photonics K.K., Hamamatsu, Japan).</p> <p>PC microscopy was performed on a Nikon Eclipse TS100-F microscope, with a Nikon CFI Achro ADL 10x/0.25 objective captured by CCD camera Jenoptik ProgRes MF.</p> <p><strong>Folder structure and file and filename description</strong><br> <br> <em>folder "source data+groundtruth"</em><br> - includes raw microscopic data <br> (uncompressed 16-bit for DIC, HMC and PC, 32-bit for QPI)<br> - includes manualy annotated groundtruth (zip file - imageJ ROI file, 1bit png mask)</p> <p>e.g. <br> DIC_01_raw.tif<br> DIC_01_groundtruth_imagejROI.zip<br> DIC_01_groundtruth_mask.png</p> <p><br> <em>folder "reconstructions"</em></p> <p>includes reconstructed images using reconstructions with highest dice coefficient achieved. </p> <p>for DIC and HMC: rDIC-Koos, rDIC-Yin, and rWeka<br> for PC: rPC-Top-Hat, rDIC-Yin, and rWeka<br> for QPI: rWeka</p> <p>note that for rWeka images numbered 01 for DIC, HMC and PC and 01-03 for QPI were used for learning.</p> <p><strong>Abbreviations</strong><br> DIC, differential image contrast<br> HMC, Hoffman modulation contrast<br> PC, phase contrast<br> QPI, quantitative phase imaging<br> rDIC-Koos, DIC/HMC image reconstruction according to Koos et al, Sci Rep. 2016;6:30420<br> rDIC-Yin, DIC/HMC image reconstruction according to Yin et al, Inf Process Med Imaging. 2011;22:384-97.<br> rPC-Yin, PC image reconstruction according to Yin et al, Med Im Anal. 2012; 16(5):1047<br> rPC-Top-Hat, Top-Hat filter according to Dewan et al, IEEE Transactions on Biomedical Circuits and<br> Systems.2014;8(5):716-728<br> rWeka, probability map using Trainable Weka segmentation according to Arganda-Carreras et al. Bioinformatics. 2017</p>
Confocal fluorescence microscopy images of the lacuno-canalicular network in bone femoral diaphysis of mice from the BionM1 project (space flight)
<p>This data set provides complementary measurements to a separate THG data set of the same study: doi: 10.5281/zenodo.1475906</p> <p>Data set for 1 sample of each of the 3 groups: Control, Space Flight and Synchro (ground control with space flight housing and feeding conditions). Contains confocal fluorescence microscopy images in tif format of 2D mosaic of selected samples and 3D stacks in selected anatomical regions of interest. See readme file for more information.</p>
Cisplatin enhances cell stiffness and decreases invasiveness rate in prostate cancer cells by actin accumulation: Confocal and atomic force microscopy
<p><strong>Summary</strong></p> <p>Dataset of imaging data related to the publication Raudenska, M., Kratochvilova, M., Vicar, T., Gumulec, J., Balvan, J., Polanska, H. Pribyl, J. & Masarik, M.:Cisplatin enhances cell stiffness and decreases invasiveness rate in prostate cancer cells by actin accumulation. <em>Scientific Reports </em><strong>2019, </strong>9, 1660</p> <p>This dataset includes image data of <em>atomic force microcopy</em> (Young modulus) and <em>confocal microscopy</em>(staining of F-actin and β-tubulin) of prostate cell lines PNT1A, 22Rv1, and PC-3. </p> <p><strong>Materials and Methods</strong></p> <p><em>Cells, cell culture conditions</em></p> <p>Cells confluent up to 50–60% were washed with a FBS-free medium and treated with a fresh medium with FBS and required antineoplastic drug concentration (IC50 concentration for the particular cell line). The cells were treated with 93 µM (PC-3), 38 µM (PNT1A), and 24 µM (22Rv1) of cisplatin (Sigma-Aldrich, St. Louis, Missouri), respectively. IC50 concentrations used for treatment with docetaxel (Sigma-Aldrich, St. Louis, Missouri) were 200nM for PC-3, 70nM for PNT1A, and 150nM for 22Rv1. </p> <p><em>Long-term zinc (II) treatment of cell cultures</em></p> <p>Cells were cultivated in the constant presence of zinc(II) ions. Concentrations of zinc(II) sulphate in the medium were increased gradually by small changes of 25 or 50 µM. The cells were cultivated at each concentration no less than one week before harvesting and their viability was checked before adding more zinc. This process was used to select zinc resistant cells naturally and to ensure better accumulation of zinc within the cells (accumulation of zinc is usually poor during the short-term treatment of prostate cancer cells). Total time of the cultivation of cell lines in the zinc(II)-containing media exceeded one year. Resulting concentrations of zinc(II) in the media (IC50 for the particular cell line) were 50 µM for the PC-3 cell line, 150 µM for the PNT1A cell line, and 400 µM for the 22Rv1 cell line. The concentrations of zinc(II) in the media and FBS were taken into account. </p> <p><em>Actin and tubulin staining</em></p> <p>β-tubulin was labeled with anti- β tubulin antibody [EPR1330] (ab108342) at a working dilution of 1/300. The secondary antibody used was Alexa Fluor® 555 donkey anti-rabbit (ab150074) at a dilution of 1/1000. Actin was labeled with Alexa Fluor™ 488 Phalloidin (A12379, Invitrogen); 1 unit per slide. For mounting Duolink® In Situ Mounting Medium with DAPI (DUO82040) was used. The cells were fixed in 3.7% paraformaldehyde and permeabilized using 0.1% Triton X-100. </p> <p><em>Confocal microscopy</em></p> <p>The microscopy of samples was performed at the Institute of Biophysics, Czech Academy of Sciences, Brno, Czech Republic. Leica DM RXA microscope (equipped with DMSTC motorized stage, Piezzo z-movement, MicroMax CCD camera, CSU-10 confocal unit and 488, 562, and 714 nm laser diodes with AOTF) was used for acquiring detailed cell images (100× oil immersion Plan Fluotar lens, NA 1.3). Total 50 Z slices was captured with Z step size 0.3 μm.</p> <p><em>Atomic force microscopy</em></p> <p>We used the bioAFM microscope JPK NanoWizard 3 (JPK, Berlin, Germany) placed on the inverted optical microscope Olympus IX‑81 (Olympus, Tokyo, Japan) equipped with the fluorescence and confocal module, thus allowing a combined experiment (AFM‑optical combined images). The maximal scanning range of the AFM microscope in X‑Y‑Z range was 100‑100‑15 µm. The typical approach/retract settings were identical with a 15 μm extend/retract length, Setpoint value of 1 nN, a pixel rate of 2048 Hz and a speed of 30 µm/s. The system operated under closed-loop control. After reaching the selected contact force, the cantilever was retracted. The retraction length of 15 μm was sufficient to overcome any adhesion between the tip and the sample and to make sure that the cantilever had been completely retracted from the sample surface. Force‑distance (FD) curve was recorded at each point of the cantilever approach/retract movement. AFM measurements were obtained at 37°C (Petri dish heater, JPK) with force measurements recorded at a pulling speed of 30 µm/s (extension time 0.5 sec).</p> <p>The Young's modulus (E) was calculated by fitting the Hertzian‑Sneddon model on the FD curves measured as force maps (64x64 points) of the region containing either a single cell or multiple cells. JPK data evaluation software was used for the batch processing of measured data. The adjustment of the cantilever position above the sample was carried out under the microscope by controlling the position of the AFM‑head by motorized stage equipped with Petri dish heater (JPK) allowing precise positioning of the sample together with a constant elevated temperature of the sample for the whole period of the experiment. Soft uncoated AFM probes HYDRA-2R-100N (Applied NanoStructures, Mountain View, CA, USA), i.e. silicon nitride cantilevers with silicon tips are used for stiffness studies because they are maximally gentle to living cells (not causing mechanical stimulation). Moreover, as compared with coated cantilevers, these probes are very stable under elevated temperatures in liquids – thus allowing long-time measurements without nonspecific changes in the measured signal.</p> <p><em>Image analysis</em></p> <p>Fluorescence microscopy data were analyzed in ImageJ 1.52h and Python 3.7.1 as follows: cells were manually segmented using actin fluorescence channel, two regions were created for analysis: whole cell and cell periphery, lining a 4 μm thick region around cell border and including most of periphery actin cytoskeleton. In these two regions following parameters were measured for both actin and tubulin fluorescence: Integrated intensity, median intensity, and following regions were measured to describe cell morphology: Cell area, Maximum caliper (max feret diameter), roundness, and aspect ratio. Moreover, stress fibers were manually segmented in every cell and following parameters were measured: number of fibers per cell, feret angle of fiber, integrated intensity, fiber length, mean intensity. Next, a standard deviation of feret angles of individual fibers was calculated relatively to mean of feret angle using a circstd function from scipy package for Python.</p> <p><strong>Identification of files</strong></p> <p><em>Microscopy data</em></p> <p>Files are separated into individual zip files. The dataset of <em>confocal microscopy </em>is separated based on treatments: untreated control, docetaxel-treated cells, cisplatin-treated cells, zinc-treated cells. Filenames actin_tubulin_Zstack_cisplatin.zip, actin_tubulin_Zstack_untreated_control.zip, actin_tubulin_Zstack_zinc.zip, actin_tubulin_Zstack_docetaxel.zip. Files included in these ZIP archives are named as follows: "cellline_treatment_FOV". Files are 3-layer 16bit tiff files with layer sequence as follows: F-Actin (Phalloidin)/b-tubulin/Hoechst 33342. The dataset contains 242 FOVs of three cell line types/three treatments + one control, files are Z-stacks made of 50 slices.</p> <p>The dataset of <em>atomic force microscopy </em>(AFM) is included in one ZIP archive "AFM_YoungModulus_SetpointHeight.zip", which includes data on Young modulus and Setpoint Height of cell lines 22Rv1, PNT1A and PC-3 and treatments zinc, docetaxel, cisplatin (+control), i.e. identical like for confocal microscopy. The file naming is as follows: "AFM_cellline_treatment_FOV_Youngmodulus.tif" for Young modulus and "AFM_cellline_treatment_FOV_setpointheight.tif" for setpoint height. The data are filtered 32-bit tiff images, where the pixel value correspond to cell stiffness (young modulus) in Pa or setpoint height in m.</p> <p><em>Confocal microscopy analysis files</em></p> <p>Following files are csv tables including image analysis of actin/tubulin staining captured by confocal microscope:</p> <p>Cytoskeleton_fluo_analysis_Cell_Cell_periphery_morphology.csv: table includes analyzed data for actin and tubulin staining in following cellular regions: cell, cell periphery. Standard ImageJ parameters regarding intensity and morphology included.</p> <p>Cytoskeleton_fluo_analysis_Fibers.csv: table includes results of manual segmentation and consequent analysis of actin stres fibers in the cells. Apart from standard ImageJ parameters, also number of stress fibers per cell and standard deviation of fiber angle relative to the cell mean angle (for details see methods) are included.</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.