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8 results for “Single Molecule Localization Microscopy”
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>
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>
Simulations of Single Molecule Localization Microscopy frames with scattered single emitters
<p>Datasets used in the work "Combining deep learning with SUPPOSe and Compressed Sensing for SNR-enhanced localization of overlapping emitters".</p> <p>The file <strong>Sample dataset.zip</strong> contains simulated images of frames of Single-Molecule Localization Microscopy. In the directory structure, Q is the number of emitters, d is the emitter distance (in pixels), imax is the image maximum intensity and i indexes different noise realizations. There are three images within each folder: <strong>X</strong> is a noiseless image, <strong>Y</strong> is the image with noise and background and <strong>Z</strong> is a denoised image predicted using a convolutional neural network.</p> <p>The file <strong>Train dataset.zip</strong> contains 5000 simulated pairs of images of single emitters distributed randomly that were used to train a convolutional neural network for denoising. The folder <strong>X</strong> contains noiseless images and the folder <strong>Y</strong> contains the corresponding images with noise and background.</p> <p>In all cases, a Gaussian PSF with size <span class="math-tex">\(\sigma = 3\)</span> px was used as the PSF of the imaging system, Noise is modeled as a Poisson process with a dark signal <span class="math-tex">\(i_{dark} = 10\)</span>.</p> <p>Corresponding author: Axel M. Lacapmesure (alacapmesure@fi.uba.ar)</p> <p> </p> <p><strong>CHANGELIST</strong></p> <ul> <li>Version 2: corrected all file extensions in "Train dataset.zip" that were wrong.</li> </ul>
Label-free adaptive optics single-molecule localization microscopy for whole zebrafish
<p>The specimen-induced aberration has been a major factor limiting the imaging depth of single-molecule localization microscopy (SMLM). Here, we report the application of label-free wavefront sensing adaptive optics to SMLM for deep-tissue super-resolution imaging. The proposed system measures complex tissue aberrations from intrinsic reflectance rather than fluorescence emission and physically corrects the wavefront distortion more than three-fold stronger than the previous limit. This enables us to resolve sub-diffraction morphologies of cilia and oligodendrocytes in whole zebrafish as well as dendritic spines in thick mouse brain tissues at the depth of up to 102 μm with localization number enhancement by up to 37 times and localization precision comparable to aberration-free samples. The proposed approach can expand the application range of SMLM to whole zebrafish that cause the loss of localization points owing to severe tissue aberrations.</p>
Single-molecule localization microscopy reveals the molecular organization of endogenous membrane receptors
Open the record for dataset details and reuse information.
Determination of protein stoichiometries via dual-color colocalization with single molecule localization microscopy
<p>This entry contains the datasets for the manuscript titled as listed.</p>
Correcting Artifacts in Single Molecule Localization Microscopy Analysis Arising from Pixel Quantum Efficiency Differences in sCMOS Cameras
<p>Jupyter notebooks and supplementary data for the paper "Correcting Artifacts in Single Molecule Localization<br> Microscopy Analysis Arising from Pixel Quantum Efficiency Differences in sCMOS Cameras".</p>
Enabling single-molecule localization microscopy in turbid food emulsions
<p>Supplementary and raw data</p>
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