Leica Science Lab - Topic : Fluorescence Microscopy https://www.leica-microsystems.com//solutions/life-science/fluorescence/?tx_leicaacademy_pi2%5Baction%5D=listPagesForTopic&tx_leicaacademy_pi2%5Bcontroller%5D=Topic&tx_leicaacademy_pi2%5Btopic%5D=21&cHash=02519c6e89d6e0bb8fbe5e5b6db54e80 Article tagged with Fluorescence Microscopy en-US https://www.leica-microsystems.com/74564 Fluorescence Microscopy Role of Mucins and Glycosylation in Dry Eye Disease This article shows how fast, high-contrast, and sharp imaging of stratified human corneal epithelial cells with THUNDER imaging technology for dry eye disease (DED) research allows membrane ridges to be resolved. DED is common and can greatly affect the patient’s quality of life for a prolonged time. The mucin proteins tethered on corneal epithelial cell membranes are hypothesized to provide barrier integrity and lubricate the eye. How mucins and their glycosylation factor into the pathogenesis of DED is not well understood due to a lack of direct and high-speed visualization methods. Sharp, high-contrast imaging of mucin layers has proven challenging with conventional widefield microscopy. However, this study demonstrates that membrane-tethered mucins expressed by corneal epithelial cells can be imaged with high-contrast using a THUNDER Imager Tissue and Large Volume Computational Clearing (LVCC). https://www.leica-microsystems.com/science-lab/role-of-mucins-and-glycosylation-in-dry-eye-disease/ Fri, 10 Sep 2021 11:56:00 +0000 BSc Chunzi Liu, PhD Olga Davydenko, PhD Cristina Rohena, PhD David R. Barbero https://www.leica-microsystems.com/73416 Fluorescence Microscopy Skeletal Muscle Adaptation and Fibrotic Diseases The mechanisms of how skeletal muscle adapts to fibrotic pathologies can be investigated more efficiently with fast, high-contrast imaging of C2C12 mouse myoblast cells which is described in this article. A better understanding of muscle fibrotic diseases and injured cell regeneration may lead to the development of novel therapies. Muscle regeneration studies require imaging solutions which can quickly screen muscle tissues and accurately assess cell damage. For this study, myoblast cells were labelled with a DNA-damage marker and then fluorescence imaging was done using a THUNDER Imager Live Cell with Large Volume Computational Clearing (LVCC). The quantification of DNA-marker foci, i.e., cell damage, was done more easily compared to conventional widefield microscopy. https://www.leica-microsystems.com/science-lab/skeletal-muscle-adaptation-and-fibrotic-diseases/ Thu, 02 Sep 2021 10:33:00 +0000 PhD Jen-Yi Lee, PhD James DeRose, PhD David R. Barbero https://www.leica-microsystems.com/74354 Confocal Microscopy Fluorescence Microscopy Spectroscopic Evaluation of Red Blood Cells Hemoglobinopathies are a major healthcare problem. This study presents a possible diagnostic tool for thalassemia which is based on confocal spectroscopy. This approach exploits spectral detection and white light laser excitation to acquire autofluorescence signals from red blood cells (RBCs). https://www.leica-microsystems.com/science-lab/spectroscopic-evaluation-of-red-blood-cells/ Mon, 30 Aug 2021 08:07:00 +0000 Dr. Giulia Ossato https://www.leica-microsystems.com/73674 Neuroscience Fluorescence Microscopy Cortical Contributions to Complex Learning This article describes the use of sharp, high contrast imaging to facilitate assessment of viral-construct placement in rat cortical brain tissue. Complex learning is often investigated through use of a rodent model. To avoid the limitations associated with permanent rat brain lesions, chemogenetic-induced temporary inactivations can be conducted. This approach involves using a viral construct to deliver foreign DNA (a receptor) and a fluorescent marker to cells within specific brain regions. https://www.leica-microsystems.com/science-lab/cortical-contributions-to-complex-learning/ Tue, 17 Aug 2021 11:53:00 +0000 PhD James DeRose https://www.leica-microsystems.com/73390 Super-Resolution Fluorescence Microscopy Confocal Microscopy Visualization of Submitochondrial Protein Distributions By allowing visualization of submitochondrial protein distributions, fluorescence nanoscopy offers significant advantages for understanding cell death control. How cells manage, and control mitochondrial intermembrane proteins involved in apoptosis, such as Cytochrome c and DIABLO/SMAC is not completely defined. In this study, we used two-color STED nanoscopy to gain insight into the submitochondrial distribution of Cytochrome c and DIABLO/SMAC, two apoptotic effectors. https://www.leica-microsystems.com/science-lab/visualization-of-submitochondrial-protein-distributions/ Mon, 16 Aug 2021 09:32:00 +0000 https://www.leica-microsystems.com/73407 Super-Resolution Fluorescence Microscopy Confocal Microscopy DNA Replication in Cancer Cells DNA synthesis can be impeded by collisions between the DNA replication machinery and co-transcriptional R-loops leading to a major source of genomic instability in cancer cells. In this paper we showed that the ATP-dependent chromatin remodelling INO80 complex promotes resolution of R-loops to prevent replication-associated DNA damage in cancer cells. We use STED super resolution nanoscopy to measure with unprecedented precision colocalization between INO80 and R-loops. https://www.leica-microsystems.com/science-lab/dna-replication-in-cancer-cells/ Tue, 27 Jul 2021 10:11:00 +0000 https://www.leica-microsystems.com/72956 Fluorescence Microscopy Widefield Microscopy Monitoring Immunosuppressive Mechanisms from Infection This application note discusses the importance of fast, sensitive, and haze-free imaging for the monitoring of immunosuppression in mouse lung epithelial tissue infected with the Puerto Rico 8 influenza virus. The study of how basal cell inhibition and immunosuppressive mechanism activation occur in virally infected lung tissue is critical for a better understanding of how viruses cause lung injury. https://www.leica-microsystems.com/science-lab/monitoring-immunosuppressive-mechanisms-from-infection/ Tue, 29 Jun 2021 09:42:00 +0000 PhD James DeRose, PhD David R. Barbero https://www.leica-microsystems.com/72687 Fluorescence Microscopy Widefield Microscopy “Brains-In-A-Dish” from Induced Pluripotent Stem Cells (iPSCs) This article discusses the benefits of using the THUNDER technology for imaging inside 3D human cortical brain organoids. These organoids are derived from human induced pluripotent stem cells (iPSCs) and can act as functional 3D “brains-in-a-dish” which can be used as models to study glia cell development and disorders like autism spectrum disorders. https://www.leica-microsystems.com/science-lab/brains-in-a-dish-from-induced-pluripotent-stem-cells-ipscs/ Fri, 28 May 2021 08:57:00 +0000 Samantha Lanjewar, Steven Sloan, PhD James DeRose, Christopher Murphy, PhD David R. Barbero https://www.leica-microsystems.com/72745 Widefield Microscopy Fluorescence Microscopy Fission Yeast Actomyosin Ring Integrity This article demonstrates that the stress-activated protein kinase pathway (SAPK) and its effector, MAPK Sty1, downregulates CAR assembly in the fission yeast Schizosaccharomyces pombe when its integrity becomes compromised during cytoskeletal damage and stress by the reduction of formin For3 levels. Cytokinesis, which enables the physical separation of daughter cells once mitosis has been completed, is executed in fungal and animal cells by a contractile actin- and myosin-based ring (CAR). https://www.leica-microsystems.com/science-lab/fission-yeast-actomyosin-ring-integrity/ Fri, 28 May 2021 07:56:00 +0000 PhD James DeRose https://www.leica-microsystems.com/72677 Widefield Microscopy Fluorescence Microscopy Developing Heart Pacemaker Cells from Cardiac Spheroids During the last decade, 3D cell culture has been established as a more realistic model compared to classical 2D culture systems. Cells can develop into miniature 3D objects, so called spheroids, which resemble organs in function and development better than 2D cell cultures. This fact makes them a great tool for the study of diseases in vitro. In addition, it is imaginable to use 3D cell culture to produce “spare parts” which ultimately can be implanted into a living organism for the sake of curing disease. https://www.leica-microsystems.com/science-lab/developing-heart-pacemaker-cells-from-cardiac-spheroids/ Wed, 26 May 2021 08:04:00 +0000 Dr. Christoph Greb, PhD James DeRose https://www.leica-microsystems.com/72461 Widefield Microscopy Fluorescence Microscopy Into the Third Dimension with "Wow Effect"- Observe Cells in 3D and Real-Time Life is fast, especially for a cell. As a rule, cells should be examined under physiological conditions which are as close as possible to their natural environment. New technologies offer tremendous performance from camera-based fluorescence systems that operate at full resolution in one shot. This article describes how new technologies are being used to effectively remove unwanted image content from areas outside the focal plane in real time. It is argued that these new approaches and the exchange of data are driving progress in science. https://www.leica-microsystems.com/science-lab/into-the-third-dimension-with-wow-effect-observe-cells-in-3d-and-real-time/ Wed, 19 May 2021 12:17:00 +0000 https://www.leica-microsystems.com/72456 Widefield Microscopy Fluorescence Microscopy Observing 3D Cell Cultures During Development 3D cell cultures, such as organoids and spheroids, give insights into cells and their interactions with their microenvironment. These 3D cell cultures are playing an increasingly important role for researchers investigating novel cancer therapies, drugs to cure Alzheimer’s disease, or personalized microfluidics to study chemotherapeutic efficiency in cancer patients. With a new imaging system, it is now easier to examine 3D cell cultures during development in real time at the cell level. For the microscopic imaging of three-dimensional samples, various options are available, such as confocal or light sheet microscopy. https://www.leica-microsystems.com/science-lab/observing-3d-cell-cultures-during-development/ Wed, 19 May 2021 09:23:00 +0000 https://www.leica-microsystems.com/72481 Widefield Microscopy Fluorescence Microscopy Cytosolic Calcium Ions in Melanoma Cancer Cells In this article, a ratiometric fluorometric method for cytosolic calcium ion (Ca2+) measurement in cultured melanoma cells using Fura 2-AM cell loading and fluorescence microscopy imaging is presented in detail. Cytosolic Ca2+ levels are maintained at low nanomolar concentrations and disruption of Ca2+ homeostasis is associated with cell/tissue damage. https://www.leica-microsystems.com/science-lab/cytosolic-calcium-ion-measurements/ Wed, 12 May 2021 07:56:00 +0000 PhD James DeRose https://www.leica-microsystems.com/72285 Widefield Microscopy Fluorescence Microscopy Live-Cell Imaging Download The Guide to Live Cell Imaging In life science research, live cell imaging is an indispensable tool to visualize cells in a state as in vivo as possible. This E-book reviews a wide range of important considerations to take to ensure successful live cell imaging. https://www.leica-microsystems.com/science-lab/e-book-the-guide-to-live-cell-imaging/ Wed, 05 May 2021 16:15:00 +0000 https://www.leica-microsystems.com/48987 Fluorescence Microscopy Widefield Microscopy Studying Cell Division Cell division is a biological process during which all cellular components must be distributed among the daughter cells. The division process requires firm coordination for success. Microscopy is utilized to observe this procedure in living cells. https://www.leica-microsystems.com/science-lab/studying-cell-division/ Tue, 09 Mar 2021 13:25:00 +0000 Ph.D. Javier Encinar del Dedo, Ph.D. Elena Rebollo, Dr. Christoph Greb, PhD James DeRose https://www.leica-microsystems.com/71300 Fluorescence Microscopy Widefield Microscopy Viral Infections – Studying Influenza-host Interactions in 3D Specimens Stefan Finke studies virus-host interactions. It turns out that the ways the virus and host interact are different when comparing results from classical laboratory monolayer cell lines and “close to in vivo” infection models. https://www.leica-microsystems.com/science-lab/viral-infections-studying-influenza-host-interactions-in-3d-specimens/ Thu, 04 Mar 2021 08:32:00 +0000 Dr. Christoph Greb https://www.leica-microsystems.com/48471 Fluorescence Microscopy Widefield Microscopy Finding new Scaffolds for Tissue Engineering Tissue engineers use biomaterials for a variety of applications from drug delivery to supporting the regeneration of damaged or lost tissues to creating in vitro disease models. Scaffold architecture can be tailored to specific tissue engineering applications. Characterizing scaffold morphology and porosity through imaging is crucial to the fabrication of modular biomaterials. Widefield microscopy can reach its limits when used for tissue engineering due to the thickness and optical characteristics of the tissue. THUNDER Imagers equipped with the Computational Clearing technology offer the advantages of widefield microscopy, but overcome the out-of-focus blur or “haze” typical when imaging thick, 3D specimens. https://www.leica-microsystems.com/finding-new-scaffolds-for-tissue-engineering/ Wed, 17 Feb 2021 09:26:00 +0000 Mollie Smoak, Dr. Christoph Greb, PhD James DeRose https://www.leica-microsystems.com/48638 Widefield Microscopy Fluorescence Microscopy The Power of Pairing Adaptive Deconvolution with Computational Clearing Deconvolution is a computational method used to restore the image of the object that is corrupted by the point spread function (PSF) along with sources of noise. In this technical brief, learn how the deconvolution algorithm offered by Leica Microsystems allows you to overcome losses in image resolution and contrast in widefield (WF) fluorescence microscopy due to the wave nature of light and the diffraction of light by optical elements. Explore the methods of user-controlled or automated deconvolution to see and resolve more structural detail. https://www.leica-microsystems.com/the-power-of-pairing-adaptive-deconvolution-with-computational-clearing/ Tue, 16 Feb 2021 11:22:00 +0000 PhD Vikram Kohli, PhD James M. Marr, Dr. Oliver Schlicker, Dr. Levi Felts