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175 results for “single-molecule”
Datasets for "Single-molecule and super-resolved imaging deciphers membrane behaviour of onco-immunogenic CCR5"
<p><strong>Flow cytometry</strong></p> <p>Modality / instrument: <em>Flow cytometer</em> <em>(CytoFLEX LX, Beckman Coulter)</em></p> <p>File format:<em> FCS + XIT (CytExpert, Beckman Coulter).</em></p> <p>Samples and acquisitions:</p> <p>Fluorescent fusions in live Chinese Hamster ovary (CHO) cells.</p> <table> <tbody> <tr> <td> <p><em>File</em></p> </td> <td> <p><em>Cell line</em></p> </td> <td> <p><em>Runs</em></p> </td> <td> <p><em>Cells counted</em></p> </td> </tr> <tr> <td> <p>CONTROL.fcs</p> </td> <td> <p>CHO wild-type</p> </td> <td> <p>1</p> </td> <td> <p>7000</p> </td> </tr> <tr> <td> <p>GFP-CCR5.fcs</p> </td> <td> <p>CHO-GFP-CCR5</p> </td> <td> <p>1</p> </td> <td> <p>7000</p> </td> </tr> <tr> <td> <p>Exp_20220916_1_GFP.xit</p> </td> <td> <p>N/A - metadata</p> </td> </tr> </tbody> </table> <p>Approx. size 6 MB</p> <p> </p> <p><strong>PaTCH microscopy images</strong></p> <p>Imaging modality / instrument: <em>Brightfield</em> + <em>PaTCH fluorescence microscopy</em></p> <p>Image format:<em> OME TIFF (16 bit) + MicroManager metadata files</em></p> <p>Microscope settings:</p> <p><em>488 nm triggered excitation; split red/green detection, cropped to green (GFP) channel only; 10 ms/frame laser exposure; 13.5 ms/frame-to-frame; 53 nm/px. Photometrics Prime95b CMOS.</em></p> <p>Samples and acquisitions:</p> <p>Fluorescent fusions of GFP-CCR5 receptor in live CHO cells imaged with and without 100 nM CCL5 ligand. Each subfolder corresponds to a field of view and contains one brightfield and one PaTCH acquisition of the same cell.</p> <table> <tbody> <tr> <td> <p>Folder</p> </td> <td> <p>Condition</p> </td> <td> <p>Fields of view</p> </td> </tr> <tr> <td> <p>AC6 CONTROL sc</p> </td> <td> <p>CCL5-</p> </td> <td> <p>11</p> </td> </tr> <tr> <td> <p>AC6 CCL5 sc</p> </td> <td> <p>CCL5+ (100 nM)</p> </td> <td> <p>10</p> </td> </tr> </tbody> </table> <p>Approx. size before compression: 14 GB</p> <p> </p> <p><strong>Structured illumination microscopy - volumetric stacks</strong></p> <p>Imaging modality / instrument: <em>SIM fluorescence microscopy (custom setup at NPL based on Olympus IX71)</em></p> <p>Image format:<em> OME TIFF (16 bit) with intrinsic metadata (voxel size)</em></p> <p>Microscope settings: <em>638 nm excitation; 60x/1.3 NA; Flash 4.0, Hamamatsu Photonics. For additional details see the reference below (Hunter et al, bioRxiv).</em></p> <p>Samples and acquisitions:</p> <p>Dylight 650-MC-5 labeled CCR5 receptor in fixed CHO-CCR5 cells, imaged with and without 100 nM CCL5 ligand. Each acquisition is of a unique field of view and contains one SIM reconstruction as an XYZ volumetric stack. ‘Basal membrane’ acquisitions consist of 5 slices at 200 nm z-intervals across the range of the basal membrane. ‘Whole cell' acquisitions are made up of 7 slices with 500 nm z-interval ranging from just below the basal membrane to just above the apical membrane. </p> <table> <tbody> <tr> <td>Folder</td> <td>Subfolder/condition</td> <td>Fields of view</td> </tr> <tr> <td>Basal membrane</td> <td>CCL5-</td> <td>5</td> </tr> <tr> <td>Basal membrane</td> <td>CCL5+ (100 nM)</td> <td>6</td> </tr> <tr> <td>Whole cells</td> <td>CCL5-</td> <td>5</td> </tr> <tr> <td>Whole cells</td> <td>CCL5+ (100 nM)</td> <td>8</td> </tr> </tbody> </table> <p>Approx. size before compression: 300 MB</p>
Single-molecule DNA methylation patterns of full-length human-specific LINE-1 (L1HS) retrotransposons in a panel of cell lines.
<p>We used bs-ATLAS-seq to comprehensively map the genomic location and assess the DNA methylation status of full-length human-specific LINE-1 elements (L1HS). The approach capture region 1-210 of L1HS elements, which corresponds to the most 5' end of its promoter sequence. This was performed in a panel of 12 human primary or transformed cell lines (BJ, IMR90, MRC5, H1, K562, HCT116, HeLa S3, HepG2, MCF7, HEK-293, HEK-293T, 2102Ep), many being shared with the encode project.</p> <p>These datasets provide a visualization for DNA methylation patterns at the single molecule level for each L1HS loci.</p>
Super-Resolved FRET Imaging by Confocal Fluorescence-Lifetime Single-Molecule Localization Microscopy
<p>FRET-based methods are a special tool for detecting interactions between (bio)molecules and their immediate environment. The spatial distribution of molecular interactions and functional states can be seen using FLIM (Fluorescence Lifetime IMaging) and FRET imaging. The spatial information, accuracy, and dynamic range of the observed signals are, however, constrained by the fact that conventional FLIM and FRET imaging only provides average information over an ensemble of molecules within a diffraction-limited volume. On the other hand, conventional Single Molecule Localization Microscopy (SMLM) relies on highly sensitive multi-pixel detectors (e.g. sCMOS or EM-CCD) whose time resolution is not suitable for fluorescence lifetime measurements.</p> <p>Here, we demonstrate a method for obtaining super-resolved FRET imaging using confocal fluorescence-lifetime single-molecule localization microscopy. The proof of concept was carried out using a DNA origami sample for performing DNA-PAINT measurements in combination with fluorogenic probes for reducing background signal. With this method, We show that FRET events separated by sub-diffraction distances can be distinguished based on lifetime modifications.</p>
Addressable Nanoantennas with Cleared Hotspots for Single-Molecule Detection on a Portable Smartphone Microscope
<p>The advent of highly sensitive photodetectors and the development of photostabilization strategies made detecting the fluorescence of single molecules a routine task in many labs around the world. However, to this day, this process requires cost-intensive optical instruments due to the truly nanoscopic signal of a single emitter. Simplifying single-molecule detection would enable many exciting applications, <em>e.g.</em> in point-of-care diagnostic settings, where costly equipment would be prohibitive. Here, we introduce addressable NanoAntennas with Cleared HOtSpots (NACHOS) that are scaffolded by DNA origami nanostructures and can be specifically tailored for the incorporation of bioassays. Single emitters placed in the NACHOS emit up to 461-fold (average of 89±7-fold) brighter enabling their detection with a customary smartphone camera and an 8-US-dollar objective lens. To prove the applicability of our system, we built a portable, battery-powered smartphone microscope and successfully carried out an exemplary single-molecule detection assay for DNA specific to antibiotic-resistant <em>Klebsiella pneumonia</em> „on the road “. Here we demonstrate the raw data on which our findings based on.</p>
Widespread Polycistronic Transcripts in Fungi Revealed by Single-Molecule mRNA Sequencing
<p>Genes in prokaryotic genomes are often arranged into clusters and co-transcribed into poly- cistronic RNAs. Isolated examples of polycistronic RNAs were also reported in some higher eukaryotes but their presence was generally considered rare. Here we developed a long- read sequencing strategy to identify polycistronic transcripts in several mushroom forming fungal species including Plicaturopsis crispa, Phanerochaete chrysosporium, Trametes ver- sicolor, and Gloeophyllum trabeum. We found genome-wide prevalence of polycistronic transcription in these Agaricomycetes, involving up to 8% of the transcribed genes. Unlike polycistronic mRNAs in prokaryotes, these co-transcribed genes are also independently transcribed. We show that polycistronic transcription may interfere with expression of the downstream tandem gene. Further comparative genomic analysis indicates that polycis- tronic transcription is conserved among a wide range of mushroom forming fungi. In sum- mary, our study revealed, for the first time, the genome prevalence of polycistronic transcription in a phylogenetic range of higher fungi. Furthermore, we systematically show that our long-read sequencing approach and combined bioinformatics pipeline is a generic powerful tool for precise characterization of complex transcriptomes that enables identifica- tion of mRNA isoforms not recovered via short-read assembly.</p>
Data and code for "Tweezepy: A Python package for calibrating forces in single-molecule video-tracking instruments"
<p>Data and code for "Tweezepy: A Python package for calibrating forces in single-molecule video-tracking instruments."</p> <p>Data includes representative real and simulated bead trajectories used in the manuscript.</p> <p>Code includes all simulations, analysis, and plot details for the Figures in the manuscript. </p> <p>See included README.txt for more details.</p>
Dynamic FRET example videos related to "Mars, a molecule archive suite for reproducible analysis and reporting of single-molecule properties from bioimages"
<p>Videos of dynamic switching between iso-I and iso-II conformations of a holiday junction at 50 mM Magnesium resulting in high and low FRET from Cy3 and Alexa647 labels positioned on the arms. Holiday junctions are surface immobilized through a biotin attachment and imaged using TIRF microscopy. The camera sensor is split using a dual view so that the acceptor emission is on the top and the donor emission is on the bottom. Videos from each position are provided as compressed zip files containing a sequence of tif files and associated metadata text file. Image sequences were collected using Micro-Manager 2.0 using ALEX or alternating laser excitation with alternating 637 and 532 pulses separated as two different channels. Beam profile images are provided for 637 and 532 excitation allowing for correction of the non-uniform beam profiles. The following 2D affine transformation matrix can be used to transform from the top acceptor emission region to the bottom donor emission region during processing.</p> <p>Affine 2D transformation from top to bottom: (m00, m01, m02, m10, m11, m12), (1.00276, 0.000208, 1.01236, 0.000267, 1.00312, 507.21025)</p> <p>A detailed image processing workflow for this dataset using Mars can be found under the example section at <a href="https://duderstadt-lab.github.io/mars-docs/">https://duderstadt-lab.github.io/mars-docs/</a> or directly at <a href="https://duderstadt-lab.github.io/mars-docs/examples/FRET_dynamic/">https://duderstadt-lab.github.io/mars-docs/examples/FRET_dynamic/</a></p>
Single-molecule FRET reveals multiscale chromatin dynamics modulated by HP1α-Fig. 2def
<p>smTIRF-FRET Data for Fig 2, for "Single-molecule FRET reveals multiscale chromatin dynamics modulated by HP1α"</p>
Single-molecule FRET reveals multiscale chromatin dynamics modulated by HP1α-Fig. 7cde
<p>smTIRF-FRET Data for Fig 7, for "Single-molecule FRET reveals multiscale chromatin dynamics modulated by HP1α"</p>
Dataset supporting the paper "Bipolar single-molecule electroluminescence and electrofluorochromism. Physical Review Research 5, 033027 (2023)"
<p>Dataset corresponding to theoretical calculations in the paper "Bipolar single-molecule electroluminescence and electrofluorochromism. Physical Review Research 5, 033027 (2023)" DOI: https://doi.org/10.1103/PhysRevResearch.5.033027</p> <p>Please cite as:</p> <p>Tzu-Chao Hung, Roberto Robles, Brian Kiraly, Julian H. Strik, Bram A. Rutten, Alexander A. Khajetoorians, Nicolas Lorente and Daniel Wegner. Dataset supporting the paper "Bipolar single-molecule electroluminescence and electrofluorochromism. Physical Review Research 5, 033027 (2023)" DOI:10.5281/zenodo.13768737</p> <p>List of files:</p> <p>Several folders corresponding to the figures of the paper. They contain:</p> <p>CONTCAR files: relaxed structures in VASP format. They can be visualized with VESTA (https://jp-minerals.org/vesta/en/).</p> <p>.agr: grace files (https://plasma-gate.weizmann.ac.il/Grace/).</p> <p>Image files in png format.</p>
A survey of the sorghum transcriptome using single-molecule long reads
<p>Alternative splicing and alternative polyadenylation (APA) of pre-mRNAs greatly contribute to transcriptome diversity, coding capacity of a genome and gene regulatory mechanisms in eukaryotes. Second-generation sequencing technologies have been extensively used to analyze transcriptomes. However, a major limitation of short-read data is that it is difficult to accurately predict full-length splice isoforms. Here we sequenced the sorghum transcriptome using Pacific Biosciences single molecule real time long-read isoform sequencing and developed a pipeline called TAPIS (Transcriptome Analysis Pipeline for Isoform Sequencing) to identify full-length splice isoforms and APA sites. Our analysis reveals transcriptome-wide full-length isoforms at an unprecedented scale with over 11,000 novel splice isoforms. Additionally, we uncover APA of ~11,000 expressed genes and more than 2,100 novel genes. These results greatly enhance sorghum gene annotations and aid in studying gene regulation in this important bioenergy crop. The TAPIS pipeline will serve as a useful tool to analyze Iso-Seq data from any organism.</p>
Multi-cell type deconvolution using a probabilistic model for single-molecule DNA methylation haplotypes
<p>Files required to run deconvolution with CelFIE-ISH and Epistate, in U250 regions from Loyfer et al. 2023, in both "pat" and "epiread" formats. </p>
Reference data and analysis software for "Four-color single-molecule imaging with engineered tags resolves the molecular architecture of signaling complexes in the plasma membrane"
<p>Reference data set for the single molecule co-tracking analysis presented in "Four-color single-molecule imaging with engineered tags resolves the molecular architecture of signaling complexes in the plasma membrane". Corresponding author for further inquiries:</p> <p>Prof. Dr. Jacob Piehler</p> <p>University of Osnabrück, Department of Biology/Chemistry, Division of Biophysics, Barbarastr. 11, 49076 Osnabrück, Germany</p> <p>https://www.biophysik.uni-osnabrueck.de/</p>
SupportingDataset Identification and Quantification of Within-Burst Dynamics in Singly-Labeled Single-Molecule Fluorescence Lifetime Experiments
<p>The Jupyter notebooks and resulting files used to demonstrate divisor-based mpH<sup>2</sup>MM. The analysis is demonstrated with both simulations and analyses of alpha-synuclein.</p> <ol> <li> <p><em><strong>Notebooks.zip</strong></em>:* Zip file containing the Jupyter notebooks for producing, analyzing and visualizing the simulated photon trajectories. Note: this folder contains all code needed to reproduce simulations. All other files related to the simulations are produced by one of the notebooks in this trajectory. However, as simulations can take a long time, the various results files are included in this repository so that notebooks can be run from intermediate steps.</p> <ol> <li> <p><strong>1-PIFE-pybromo-sims.ipynb</strong> : The code for producing simulated diffusion trajectories and photon-HDF5 files of two-state systems undergoing transition dynamics (the results of this notebook are stored in the sub-folder <em>PyBroMo_photonHDF5</em>)</p> </li> <li> <p><strong>2-PIFE-mpH2MM-sim-[lifetime components].ipynb </strong>: Notebooks performing divisor-based mpH<sup>2</sup>MM on simulated datasets for a given combination of lifetime states. (these notebooks store files that are contained in the sub-folder <em>H2MMresults</em>)</p> </li> <li> <p><strong>3-PIFE-mpH2MM-compiled-plots.ipynb</strong> : Jupyter notebook for producing figures comparing all results globally</p> </li> <li> <p><strong>532nm_IRF_19-10-2021.csb</strong>: the file containing the experimental IRF used in the simulations</p> </li> </ol> </li> <li> <p><em><strong>PyBroMo_photonHDF5.zip</strong></em>:* Zip file containing the simulated results of <em>1-PIFE-pybromo-sims</em> notebook as photon-HDF5 files (1 file per transition rate/lifetime combination)</p> </li> <li> <p><strong>PIFE-sim-dynamicmix_[lifetime components]_result.hdf5</strong>: special HDF5 files containing the results of each notebook in <em>Notebooks</em>, which are used by <em>3-PIFE-mpH2MM-compiled-plots</em></p> </li> <li> <p><strong>PIFE-mpH2MM-alpha-syn-vFinal.ipynb</strong>: divisor-based mpH<sup>2</sup>MM analysis of alpha-synuclein smPIFE data</p> </li> <li> <p><strong>H2MM-Lifetime_example.ipynb</strong>: A demonstration of divisor-based mpH<sup>2</sup>MM using nsALEX-smFRET data. This method could potentially demonstrate states differentiated in lifetimes independently of potential changes in E & S.</p> </li> <li> <p><strong>Template_ltH2MM.ipynb</strong>: An easy-to-follow implementation of divisor-based mpH<sup>2</sup>MM demonstrated on a single alpha-synuclein experimental data acquisition file. This can be used for learning how to implement and analyze single dye fluorescence lifetime data with mpH<sup>2</sup>MM</p> </li> </ol> <p> </p> <p>* For running these notebooks, generally, all files in <em>PyBroMo_photonHDF5.zip</em> should be placed into a single directory (i.e., the files in <em>Notebooks</em>.<em>zip</em> should be placed into the same directory as the files in <em>PyBroMo_photonHDF5</em>.<em>zip</em>) as the notebooks are set to read in files from their current directory.</p>
Enabling spectrally resolved single-molecule localization microscopy at high emitter densities: Dataset
<p>The data in this dataset accompanies the various figures present in the publication 'Enabling spectrally resolved single-molecule localization microscopy at high emitter densities'. Contained are tiff files used to create the figures 2-4 and Supplementary figures 1 and 2, as well as csvs after processed with the steps described in the paper (and contained in protocol text files).</p>
Long-Term, Single-Molecule Imaging of Proteins in Live Cells with Photoregulated Fluxional Fluorophores
<p>Supporting data for paper "Long-Term, Single-Molecule Imaging of Proteins in Live Cells with Photoregulated Fluxional Fluorophores"</p>
Single-molecule FRET reveals multiscale chromatin dynamics modulated by HP1α-Fig. 1df
<p>smTIRF-FRET Data for Fig 1, for "Single-molecule FRET reveals multiscale chromatin dynamics modulated by HP1α"</p>
Nanotiming: single-molecule based, telomere-to-telomere DNA replication timing profiling by nanopore sequencing
<p>Dataset for the manuscript "Nanotiming: telomere-to-telomere DNA replication timing profiling by nanopore sequencing" by Theulot et al ,2024 (<span>https://doi.org/10.1038/s41467-024-55520-3</span>) related to the github repository (https://github.com/LacroixLaurent/NanoTiming)</p> <ul> <li>WT_rep3.tar.gz contains fast5 file from an experiment where yeast BT1 strain was grown for one doubling time with 5µM BrdU then DNA was sequenced on R9.4.1 ONT flowcell</li> <li>mod_mapping.bam contains the bam file resulting from the BrdU base calling with megalodon (v2.2.9) using our BT1 reference genome and our BrdU aware model for base-calling</li> <li>WT_rep3_nanoT.bed.gz contains the reads coordinates from the mod_mappings file</li> <li>WT_rep3_nanoT_alldata.rds contains the BrdU profiles for each reads of the mod_mappings file, with the BrdU signal binned in 1kb non overlaping windows</li> <li>WT_rep3_nanoT.rds contains the genomic BrdU signal profiles by 1kb non overlaping windows</li> <li>TeloLengthDataNanoT.rds contains all the telomeric sequences extracted from the experiments reported in the Figure 4 and S19 to S23 of the manuscript with the associated filtering information and nanotiming signal.</li> </ul> <p> </p>
Datasets for "Precision and accuracy of single-molecule FRET measurements – a multi-laboratory benchmark study"
<p>Supplementary material (raw data) for Fig. 2 in "<strong>Precision and accuracy of single-molecule FRET measurements – a multi-laboratory benchmark study</strong>" to be published with Nature Methods</p> <p>The confocal data is given in ht3 and hdf5 format.</p> <p>For the TIRF data the original TIFF-stacks are uploaded including the calibration files.</p>
Benchmarking Smartphone Fluorescence-Based Microscopy with DNA Origami Nanobeads: Reducing the Gap toward Single-Molecule Sensitivity
<p>Smartphone-based fluorescence microscopy has been rapidly developing over the last few years, enabling point-of-need detection of cells, bacteria, viruses, and biomarkers. These mobile microscopy devices are cost-effective, field-portable, and easy to use, and benefit from economies of scale. Recent developments in smartphone camera technology have improved their performance, getting closer to that of lab microscopes. Here, we report the use of DNA origami nanobeads with predefined numbers of fluorophores to quantify the sensitivity of a smartphone-based fluorescence microscope in terms of the minimum number of detectable molecules per diffraction-limited spot. With the brightness of a single dye molecule as a reference, we compare the performance of color and monochrome sensors embedded in state-of-the-art smartphones. Our results show that the monochrome sensor of a smartphone can achieve better sensitivity, with a detection limit of ∼10 fluorophores per spot. The use of DNA origami nanobeads to quantify the minimum number of detectable molecules of a sensor is broadly applicable to evaluate the sensitivity of various optical instruments.</p>
ScienceDex guides
Understand access before you commit
These curated guides explain access requirements, typical timelines, costs, and reuse considerations for widely used research datasets.
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.