Leica Science Lab - Tag : HyVolution https://www.leica-microsystems.com//science-lab/tag/hyvolution/ Article tagged with HyVolution en-US https://www.leica-microsystems.com/24557 Super-Resolution Dynamic Endoplasmic Reticulum The Endoplasmic Reticulum (ER) is a dynamic, reticular or net-like structure consisting of tubules and cisternae that extend throughout the cell and occupy a large portion of the cytoplasm. In recent years, it has become clear that the ER is not a static organelle. Instead the ER is dynamic, forming contacts with nearly every intracellular membrane. Recent innovations that improve image resolution in light microscopy have advanced our ability to study the dynamics of the endoplasmic reticulum and the formation of ER contact sites with other organelles. https://www.leica-microsystems.com/science-lab/dynamic-endoplasmic-reticulum/ Fri, 30 Nov 2018 23:00:00 +0000 https://www.leica-microsystems.com/20511 Super-Resolution How to extract Image Information by Adaptive Deconvolution Confocal Laser Scanning Microscopy (CLSM) is the standard for true 3D resolved fluorescence imaging. Fast optical sectioning using flexible scanning strategies in combination with simultaneous multi-colour, high sensitivity and low noise signal detection provides maximum resolution in the spatial and temporal domain. In combination with modern approaches to image information extraction this helps the researcher to mine as much information as possible from the images acquired. Image information extraction refers to intelligent procedures for image enhancement using a priori knowledge from the imaging system. From simple glare control and optical development to intelligent and ingenious model extraction, there are many ways to see more than just the image. https://www.leica-microsystems.com/science-lab/how-to-extract-image-information-by-adaptive-deconvolution/ Tue, 18 Sep 2018 22:00:00 +0000 Dr. Jürgen Reymann https://www.leica-microsystems.com/18958 Super-Resolution The Bif-1-Dynamin 2 Membrane Fission Machinery Regulates Atg9-Containing Vesicle Generation at the Rab11-Positive Reservoirs Application example of HyVolution Super-Resolution - Atg9 is a multispanning transmembrane protein that is required for autophagosome formation. During autophagy, vesicles containing Atg9 are generated through an unknown mechanism and delivered to the autophagosome formation sites. We have previously reported that Atg9-containing membranes undergo continuous tubulation and fission during nutrient starvation in a manner dependent on the curvature-inducing protein Bif-1/Sh3glb1. Here, we identify Dynamin 2 (DNM2) as a Bif-1-interacting protein that mediates the fission of Atg9-containing membranes during autophagy. https://www.leica-microsystems.com/science-lab/the-bif-1-dynamin-2-membrane-fission-machinery-regulates-atg9-containing-vesicle-generation-at-the-rab11-positive-reservoirs/ Wed, 25 Jan 2017 13:00:00 +0000 https://www.leica-microsystems.com/18937 Super-Resolution Neuroscience Quantitative Analysis of PPT1 Interactome in Human Neuroblastoma Cells Application example of HyVolution Super-Resolution - Mutations in the CLN1 gene that encodes Palmitoyl protein thioesterase 1 (PPT1) or CLN1, cause Infantile NCL (INCL, MIM#256730). PPT1 removes long fatty acid chains such as palmitate from modified cysteine residues of proteins. The data shown here result from isolated protein complexes from PPT1-expressing SH-SY5Y stable cells that were subjected to single step affinity purification coupled to mass spectrometry (AP-MS). Prior to the MS analysis, we utilised a modified filter-aided sample preparation (FASP) protocol. Based on label free quantitative analysis of the data by SAINT, 23 PPT1 interacting partners (IP) were identified. https://www.leica-microsystems.com/science-lab/quantitative-analysis-of-ppt1-interactome-in-human-neuroblastoma-cells/ Fri, 13 Jan 2017 16:23:00 +0000 https://www.leica-microsystems.com/18925 Confocal Microscopy Super-Resolution Methods to Calibrate and Scale Axial Distances in Confocal Microscopy as a Function of Refractive Index Application example of HyVolution Super-Resolution - Accurate distance measurement in 3D confocal microscopy is important for quantitative analysis, volume visualization and image restoration. However, axial distances can be distorted by both the point spread function (PSF) and by a refractive-index mismatch between the sample and immersion liquid, which are difficult to separate. Additionally, accurate calibration of the axial distances in confocal microscopy remains cumbersome, although several high-end methods exist. In this paper we present two methods to calibrate axial distances in 3D confocal microscopy that are both accurate and easily implemented. https://www.leica-microsystems.com/science-lab/methods-to-calibrate-and-scale-axial-distances-in-confocal-microscopy-as-a-function-of-refractive-index/ Fri, 09 Dec 2016 08:29:00 +0000 https://www.leica-microsystems.com/18927 Super-Resolution Evaluation of Zebrafish as a Model to Study the Pathogenesis of the Opportunistic Pathogen Cronobacter Turicensis Application example of HyVolution Super-Resolution - Bacteria belonging to the genus Cronobacter spp. have been recognized as causative agents of life-threatening systemic infections, primarily in premature, low-birth weight and/or immune-compromised neonates. Knowledge remains scarce regarding the underlying molecular mechanisms of disease development. In this study, we evaluated the use of a zebrafish model to study the pathogenesis of Cronobacter turicensis LMG 23827T, a clinical isolate responsible for two fatal sepsis cases in neonates. https://www.leica-microsystems.com/science-lab/evaluation-of-zebrafish-as-a-model-to-study-the-pathogenesis-of-the-opportunistic-pathogen-cronobacter-turicensis/ Fri, 25 Nov 2016 14:43:00 +0000 https://www.leica-microsystems.com/18952 Super-Resolution Live-Cell Imaging Discovery of Novel Peptides Targeting Pro-Atherogenic Endothelium in Disturbed Flow Regions -Targeted siRNA Delivery to Pro-Atherogenic Endothelium in vivo Application example of HyVolution Super-Resolution - Atherosclerosis occurs preferentially in arterial regions exposed to disturbed blood flow. Targeting these pro-atherogenic regions is a potential anti-atherogenic therapeutic approach, but it has been extremely challenging. Here, using in vivo phage display approach and the partial carotid ligation model of flow-induced atherosclerosis in mouse, we identified novel peptides that specifically bind to endothelial cells (ECs) exposed to disturbed flow condition in pro-atherogenic regions. https://www.leica-microsystems.com/science-lab/discovery-of-novel-peptides-targeting-pro-atherogenic-endothelium-in-disturbed-flow-regions-targeted-sirna-delivery-to-pro-atherogenic-endothelium-in-vivo/ Thu, 10 Nov 2016 15:28:00 +0000 https://www.leica-microsystems.com/18900 Confocal Microscopy Super-Resolution P53- and Mevalonate Pathway–Driven Malignancies Require Arf6 for Metastasis and Drug Resistance Application example of HvYolution Super-Resolution - Drug resistance, metastasis, and a mesenchymal transcriptional program are central features of aggressive breast tumors. The GTPase Arf6, often overexpressed in tumors, is critical to promote epithelial–mesenchymal transition and invasiveness. The metabolic mevalonate pathway (MVP) is associated with tumor invasiveness and known to prenylate proteins, but which prenylated proteins are critical for MVP-driven cancers is unknown. We show here that MVP requires the Arf6-dependent mesenchymal program. https://www.leica-microsystems.com/science-lab/p53-and-mevalonate-pathway-driven-malignancies-require-arf6-for-metastasis-and-drug-resistance/ Wed, 19 Oct 2016 16:27:00 +0000 https://www.leica-microsystems.com/17957 Confocal Microscopy Super-Resolution Light Sheet Microscopy Neuroscience Super-Resolution Mapping of Neuronal Circuitry With an Index-Optimized Clearing Agent Super-resolution imaging deep inside tissues has been challenging, as it is extremely sensitive to light scattering and spherical aberrations. Here, we report an optimized optical clearing agent for high-resolution fluorescence imaging (SeeDB2). SeeDB2 matches the refractive indices of fixed tissues to that of immersion oil (1.518), thus minimizing both light scattering and spherical aberrations. https://www.leica-microsystems.com/science-lab/super-resolution-mapping-of-neuronal-circuitry-with-an-index-optimized-clearing-agent/ Wed, 27 Apr 2016 10:07:00 +0000 https://www.leica-microsystems.com/17673 Confocal Microscopy Super-Resolution HyVolution – Super-Resolution Imaging with a Confocal Microscope Since the invention of the microscope, there has been continual discussion about the possibility of showing more detailed features of specimens as compared to just magnifying them. In this article we describe the HyVolution concept and how the combination of confocal multiparameter fluorescence imaging at the confocal super-resolution regime with psf-based real deconvolution allows high-speed multicolor imaging with a resolution down to 140 nm. https://www.leica-microsystems.com/science-lab/hyvolution-super-resolution-imaging-with-a-confocal-microscope/ Fri, 01 Apr 2016 07:19:00 +0000 Dr. Rolf T. Borlinghaus, Dr. Constantin Kappel https://www.leica-microsystems.com/17669 Confocal Microscopy Super-Resolution HyVolution – the Smart Path to Confocal Super-Resolution Super-resolution refers to any device or method that can resolve better than the classical Abbe limit. Apart from infinite super-resolution techniques such as STED (stimulated emission depletion) and SMLM (single-molecule localization methods) that can theoretically resolve to any detail, there are also methods for limited super-resolution. Here we present HyVolution by Leica, which merges optical super-resolution and computational super-resolution. The optical part is provided by confocal microscopy, and the computational part by deconvolution. Lateral resolution of 140 nm is demonstrated. HyVolution offers multiple fluorescence recording in truly simultaneous mode. https://www.leica-microsystems.com/science-lab/hyvolution-the-smart-path-to-confocal-super-resolution/ Fri, 18 Mar 2016 14:49:00 +0000 Dr. Rolf T. Borlinghaus, Dr. Constantin Kappel https://www.leica-microsystems.com/15079 Confocal Microscopy Super-Resolution Super-Resolution – On a Heuristic Point of View About the Resolution of a Light Microscope Since super-resolution has become one of the most favored methods in biomedical research, the term has become increasingly popular. Still, there is much of confusion about what is super-resolution and what is resolution at all. Here, the classical view of microscopic resolution is discussed and some techniques that resolve better than classical are briefly introduced. The picture on the right shows the intensity distribution of an image of two points whose distance is just the Rayleigh criterion (false color coding). https://www.leica-microsystems.com/science-lab/super-resolution-on-a-heuristic-point-of-view-about-the-resolution-of-a-light-microscope/ Wed, 21 Jan 2015 16:00:00 +0000 Dr. Rolf T. Borlinghaus