Leica Science Lab - Tag : Two-Photon Imaging https://www.leica-microsystems.com//science-lab/tag/tags/two-photon-imaging/show/Tag/ Article tagged with Two-Photon Imaging en-US https://www.leica-microsystems.com/20142 Multiphoton Microscopy Confocal Microscopy High-Resolution 3D Imaging of Whole Organ after Clearing Zebrafish testis has become a powerful model for reproductive biology of teleostean fishes and other vertebrates and encompasses multiple applications in applied and basic research. Many studies have focused on 2D images, which is time consuming and implies extrapolation of results. Three-dimensional imaging of whole organs recently became an important challenge to better understand their architecture and allow cell enumeration. https://www.leica-microsystems.com//science-lab/high-resolution-3d-imaging-of-whole-organ-after-clearing/ Tue, 08 May 2018 22:00:00 +0000 https://www.leica-microsystems.com/19766 Multiphoton Microscopy Multiphoton Microscopy – a Satisfied Wish List The colorful picture shows colon tumor cells, fluorescently labelled and lineage traced from a multicolor tracer. The gray color codes for the second harmonic generation (SHG) signal from Collagen 1. Lineage traced tumor cells are shown in magenta, blue, green, yellow and red. All channels were recorded with two-photon excitation, using the SP8 DIVE by Leica Microsystems. Sample and image were kindly provided by J. van Rheenen, H. Snippert, Utrecht (the Nederlands,) and I. Steinmetz, Leica Microsystems Mannheim. https://www.leica-microsystems.com//science-lab/multiphoton-microscopy-a-satisfied-wish-list/ Wed, 25 Oct 2017 08:49:00 +0000 Dr. Rolf T. Borlinghaus, Ph.D. Bettina Griesshaber https://www.leica-microsystems.com/19707 Multiphoton Microscopy Mission Impossible Accomplished: Tunable Colors for Non-descanning Detection Leica Microsystems’ 4Tune detector, the key component of the SP8 DIVE Deep In Vivo Explorer, provides spectrally tunable image recording with non-descanning detection. An innovative solution for multiparameter multiphoton microscopy. The colorful image on the right shows multiphoton microscopy of an unstained mouse skin section acquired using the 4Tune detector. The green color codes for autofluorescence of muscle tissue. Red shows second harmonic generation of fibers upon illumination with 900 nm. The blue pattern is generated by third harmonic generation at lipid boundaries from illumination at 1230 nm. https://www.leica-microsystems.com//science-lab/mission-impossible-accomplished-tunable-colors-for-non-descanning-detection/ Wed, 23 Aug 2017 08:49:00 +0000 Dr. Rolf T. Borlinghaus, Dr. Hilmar Gugel https://www.leica-microsystems.com/19620 Multiphoton Microscopy Laser Beam Shaping for Multicolor Multiphoton Microscopy Multiphoton Microscopy is one of the current hot topics in life science research. The new Leica TCS SP8 DIVE from Leica Microsystems presents a series of beneficial new innovations, including a freely tunable non-descanning detector and an ingenious beam manipulating device VBE. The variable beam expander offers free tuning of both beam diameter and axial IR-correction for up to four IR beams simultaneously. https://www.leica-microsystems.com//science-lab/laser-beam-shaping-for-multicolor-multiphoton-microscopy/ Thu, 06 Jul 2017 08:24:00 +0000 Dr. Rolf T. Borlinghaus, Manuel Kremer https://www.leica-microsystems.com/19109 Confocal Microscopy Human NK Cell Development Requires CD56-mediated Motility and Formation of the Developmental Synapse While distinct stages of natural killer (NK) cell development have been defined, the molecular interactions that shape human NK cell maturation are poorly understood. Here we define intercellular interactions between developing NK cells and stromal cells which, through contact-dependent mechanisms, promote the generation of mature, functional human NK cells from CD34+ precursors. We show that developing NK cells undergo unique, developmental stage-specific sustained and transient interactions with developmentally supportive stromal cells, and that the relative motility of NK cells increases as they move through development in vitro and ex vivo. https://www.leica-microsystems.com//science-lab/human-nk-cell-development-requires-cd56-mediated-motility-and-formation-of-the-developmental-synapse/ Wed, 15 Feb 2017 07:36:00 +0000 Ph.D. Emily M. Mace https://www.leica-microsystems.com/18172 Multiphoton Microscopy Fluorescence Microscopy Chronic Stress in Mice Remodels Lymph Vasculature to Promote Tumour Cell Dissemination Chronic stress induces signalling from the sympathetic nervous system (SNS) and drives cancer progression, although the pathways of tumour cell dissemination are unclear. Here we show that chronic stress restructures lymphatic networks within and around tumours to provide pathways for tumour cell escape. We show that VEGFC derived from tumour cells is required for stress to induce lymphatic remodelling and that this depends on COX2 inflammatory signalling from macrophages. Pharmacological inhibition of SNS signalling blocks the effect of chronic stress on lymphatic remodelling in vivo and reduces lymphatic metastasis in preclinical cancer models and in patients with breast cancer. https://www.leica-microsystems.com//science-lab/chronic-stress-in-mice-remodels-lymph-vasculature-to-promote-tumour-cell-dissemination/ Tue, 07 Feb 2017 09:52:00 +0000 https://www.leica-microsystems.com/19049 Multiphoton Microscopy Live-Cell Imaging Confocal Microscopy Fluorescence Microscopy Neuroscience Multiphoton Microscopy Publication List Multiphoton Microscopy is an advanced technique for imaging thick samples. Applications range from the visualization of the complex architecture of the whole brain to the study of tumor development and metastasis or the responses of the immune system in living animals. On this regularly updated reference list you can find selected publications on reseach using multiphoton microscopy. https://www.leica-microsystems.com//science-lab/multiphoton-microscopy-publication-list/ Wed, 14 Dec 2016 08:44:00 +0000 Simone Diehl https://www.leica-microsystems.com/18779 Multiphoton Microscopy BABB Clearing and Imaging for High Resolution Confocal Microscopy: Counting and Sizing Kidney Cells in the 21st Century Multipohoton microscopy experiment using Leica TCS SP8 MP and Leica 20x/0.95 NA BABB immersion objective. Understanding kidney microanatomy is key to detecting and identifying early events in kidney disease. Improvements in tissue clearing and imaging have been crucial in this field, and now we report on a novel, time-efficient method to study podocyte depletion in renal glomeruli using a combination of immunofluorescence, optical clearing, confocal microscopy and 3D analysis. https://www.leica-microsystems.com//science-lab/babb-clearing-and-imaging-for-high-resolution-confocal-microscopy-counting-and-sizing-kidney-cells-in-the-21st-century/ Tue, 13 Sep 2016 09:03:00 +0000 Ph.D. Victor G. Puelles, Prof. John F. Bertram, MSc Stephen Firth, Prof. Ian Harper https://www.leica-microsystems.com/18796 Multiphoton Microscopy Confocal Microscopy Light Sheet Microscopy Neuroscience Clearing of Fixed Tissue: A Review from a Microscopist’s Perspective Chemical clearing of fixed tissues is becoming a key instrument for the three-dimensional reconstruction of macroscopic tissue portions, including entire organs. Indeed, the growing interest in this field has both triggered and been stimulated by recent advances in high-throughput microscopy and data analysis methods, which allowed imaging and management of large samples. https://www.leica-microsystems.com//science-lab/clearing-of-fixed-tissue-a-review-from-a-microscopists-perspective/ Mon, 12 Sep 2016 11:20:00 +0000 https://www.leica-microsystems.com/15645 Confocal Microscopy Live-Cell Imaging Label-free in vivo Imaging of Myelinated Axons in Health and Disease with Spectral Confocal Reflectance Microscopy We report a new technique for high-resolution in vivo imaging of myelinated axons in the brain, spinal cord and peripheral nerve that requires no fluorescent labeling. This method, based on spectral confocal reflectance microscopy (SCoRe), uses a conventional laser scanning confocal system to generate images by merging the simultaneously reflected signals from multiple lasers of different wavelengths. https://www.leica-microsystems.com//science-lab/label-free-in-vivo-imaging-of-myelinated-axons-in-health-and-disease-with-spectral-confocal-reflectance-microscopy/ Fri, 01 Jul 2016 09:29:00 +0000 https://www.leica-microsystems.com/17611 Super-Resolution Multiphoton Microscopy Live-Cell Imaging 4Pi-RESOLFT Nanoscopy Here we apply the 4Pi scheme to RESOLFT nanoscopy using two-photon absorption for the on-switching of fluorescent proteins. We show that in this combination, the lobes are so low that low-light level, 3D nanoscale imaging of living cells becomes possible. Our method thus offers robust access to densely packed, axially extended cellular regions that have been notoriously difficult to super-resolve. Our approach also entails a fluorescence read-out scheme that translates molecular sensitivity to local off-switching rates into improved signal-to-noise ratio and resolution. https://www.leica-microsystems.com//science-lab/4pi-resolft-nanoscopy/ Fri, 26 Feb 2016 12:41:00 +0000 https://www.leica-microsystems.com/14920 Multiphoton Microscopy CLEM Live-Cell Imaging Correlating Intravital Multi-Photon Microscopy to 3D Electron Microscopy of Invading Tumor Cells Using Anatomical Reference Points Cancer research unsing multiphoton microscopy and 3D electron microscopy. Correlative microscopy combines the advantages of both light and electron microscopy to enable imaging of rare and transient events at high resolution. Performing correlative microscopy in complex and bulky samples such as an entire living organism is a time-consuming and error-prone task. https://www.leica-microsystems.com//science-lab/correlating-intravital-multi-photon-microscopy-to-3d-electron-microscopy-of-invading-tumor-cells-using-anatomical-reference-points/ Fri, 24 Apr 2015 13:06:00 +0000 https://www.leica-microsystems.com/15430 Multiphoton Microscopy Confocal Microscopy Live-Cell Imaging Third Harmonic Generation Microscopy – Label-Free 3D-Tissue Imaging and Blood Flow Characterization THG microscopy as special variants of multiphoton microscopy. Third Harmonic Generation (THG) microscopy is a non-fluorescent multi-photon technique that combines the advantages of label-free imaging with restriction of signal generation to the focal spot of the scanning laser. It allows three-dimensional imaging of refraction index mismatches and of hemoglobin. https://www.leica-microsystems.com//science-lab/third-harmonic-generation-microscopy-label-free-3d-tissue-imaging-and-blood-flow-characterization/ Sat, 11 Apr 2015 17:34:00 +0000 Dr. Steffen Dietzel https://www.leica-microsystems.com/15347 Super-Resolution STED Nanoscopy: A Glimpse into the Future The well-known saying of "Seeing is believing" became even more apt in biology when stimulated emission depletion (STED) nanoscopy was introduced in 1994 by the Nobel laureate S. Hell and coworkers. This article gives an overview of the various cutting-edge implementations of STED nanoscopy and tries to shine a light into the future: imaging everything faster with unprecedented sensitivity and label-free. https://www.leica-microsystems.com//science-lab/sted-nanoscopy-a-glimpse-into-the-future/ Mon, 23 Mar 2015 18:08:00 +0000 Paolo Bianchini https://www.leica-microsystems.com/14066 Multiphoton Microscopy Live-Cell Imaging The Environment Makes the Stem Cell A recent publication in Nature shows that all stem cells divide and compete for niche space by passively "kicking out" others so that eventually one stem cell takes over the whole niche. Jacco van Rheenen and Saskia Ellenbroek talk about a new method of intravital imaging, which allows following the fate of individual stem cells over time in vivo and explains the new paradigm for stem cell development in the intestinal stem cell niche. https://www.leica-microsystems.com//science-lab/the-environment-makes-the-stem-cell/ Mon, 04 Aug 2014 16:25:00 +0000 PhD Jacco van Rheenen, PhD Saskia I. J. Ellenbroek, PhD Isabelle Köster https://www.leica-microsystems.com/14063 Confocal Microscopy Multiphoton Microscopy Smart Control for Resonant Galvo Scanners High time-resolution confocal microscopy (HTRCLSM) requires fast scanning devices. Whereas non-resonant galvo scanners allow full position control, but only at slow speed, resonant scanners allow ~25,000 lines per second, but offer much less positioning freedom. To still allow zoom and pan functions, several approaches have been tried, with varying success. The Leica confocal microscopes od the TCS series use a very smart solution that enables stepless zooming with short switching times. https://www.leica-microsystems.com//science-lab/smart-control-for-resonant-galvo-scanners/ Fri, 01 Aug 2014 08:29:00 +0000 Dr. Rolf T. Borlinghaus https://www.leica-microsystems.com/13257 Multiphoton Microscopy Live-Cell Imaging Tracking Glomerular Fate Over Long Time Distances Multi-channel multiphoton microscopy with dedicated optics for CLARITY. The glomerular filtration barrier (GFB) is a complex spatial structure within the kidney glomerulis where ultrafiltration takes place. Podocytes are critical elements of the GFB and take part in the filtration process. They have been shown to be involved in the development of kidney diseases. However, due to technical limitations, the mechanism of glomerular pathology is not well understood. https://www.leica-microsystems.com//science-lab/tracking-glomerular-fate-over-long-time-distances/ Mon, 14 Apr 2014 12:32:00 +0000 M.D., Ph.D. Janos Peti-Peterdi, PhD Isabelle Köster https://www.leica-microsystems.com/13103 Super-Resolution A New Filtering Technique for Removing Anti-Stokes Emission Background in Gated CW-STED Microscopy Stimulated emission depletion (STED) microscopy is a prominent approach of super-resolution optical microscopy, which allows cellular imaging with so far unprecedented unlimited spatial resolution. The introduction of time-gated detection in STED microscopy significantly reduces the (instantaneous) intensity required to obtain sub-diffraction spatial resolution. https://www.leica-microsystems.com//science-lab/a-new-filtering-technique-for-removing-anti-stokes-emission-background-in-gated-cw-sted-microscopy/ Tue, 25 Mar 2014 20:34:00 +0000 https://www.leica-microsystems.com/10216 Super-Resolution Multiphoton Microscopy Single-Wavelength Two-Photon Excitation-stimulated Emission Depletion (SW2PE-STED) Superresolution Imaging Two-photon microscopy, multiphoton microcopy and super-resolution imaging. We developed a new class of two-photon excitation–stimulated emission depletion (2PE-STED) optical microscope. In this work, we show the opportunity to perform superresolved fluorescence imaging, exciting and stimulating the emission of a fluorophore by means of a single wavelength. https://www.leica-microsystems.com//science-lab/single-wavelength-two-photon-excitation-stimulated-emission-depletion-sw2pe-sted-superresolution-imaging/ Fri, 23 Aug 2013 11:02:00 +0000 Paolo Bianchini https://www.leica-microsystems.com/7246 Multiphoton Microscopy Developments in Multiphoton Excitation Microscopy Basics, history, and applications of multiphoton microscopy. Honouring Goeppert-Mayer’s prediction of simultaneous two-photon absorption by an atom or molecule reported in the 1930s in her PhD dissertation thesis, we can state that multiphoton excitation (MPE) microscopy, more frequently identified with two-photon excitation (2PE) fluorescence microscopy, is a key microscopy method in many areas from medicine to biology, from biophysics to materials science. https://www.leica-microsystems.com//science-lab/developments-in-multiphoton-excitation-microscopy/ Sun, 07 Oct 2012 22:00:00 +0000 Prof. Alberto Diaspro https://www.leica-microsystems.com/6654 Multiphoton Microscopy Principles of Multiphoton Microscopy for Deep Tissue Imaging Basics of multiphoton microscopy. This interactive tutorial explains the principles of multiphoton microscopy for deep tissue imaging. Multiphoton microscopy uses excitation wavelengths in the infrared taking advantage of the reduced scattering of longer wavelengths. This makes multiphoton imaging the perfect tool for deep tissue imaging in thick sections and living animals. https://www.leica-microsystems.com//science-lab/principles-of-multiphoton-microscopy-for-deep-tissue-imaging/ Thu, 23 Aug 2012 22:00:00 +0000 Dr. Andrea Mülter https://www.leica-microsystems.com/5901 Coherent Raman Scattering (CRS) CARS Microscopy: Imaging Characteristic Vibrational Contrast of Molecules Coherent anti-Stokes Raman scattering (CARS) microscopy is a technique that generates images based on the vibrational signatures of molecules. This imaging methods does not require labeling, yet molecular specific information can be obtained from a range of important bio-molecular compounds. https://www.leica-microsystems.com//science-lab/cars-microscopy-imaging-characteristic-vibrational-contrast-of-molecules/ Mon, 16 Jul 2012 22:00:00 +0000 Ph.D. Eric Olaf Potma https://www.leica-microsystems.com/5929 Multiphoton Microscopy Confocal Microscopy Order versus Disorder In modern biomaterial design the generation of an environment mimicking some of the extracellular matrix features is envisaged to support molecular cross-talk between cells and scaffolds during tissue formation/remodeling. In bone substitutes chemical biomimesis has been particularly exploited; conversely, the relevance of pre-determined scaffold architecture for regenerated bone outputs is still unclear. https://www.leica-microsystems.com//science-lab/order-versus-disorder/ Thu, 19 Apr 2012 22:00:00 +0000 Paolo Giannoni, Paolo Bianchini, Monica Sandri, Roberto Marotta, Giuseppe Firpo, Ugo Valbusa, Anna Tampieri, Prof. Alberto Diaspro, Paolo Bianco, Rodolfo Quarto https://www.leica-microsystems.com/4713 Fluorescence Microscopy Modern Fluorescent Proteins and their Biological Applications Here we present two review articles on fluorescent proteins and their biological applications. These first article reviews our current knowledge of blue, green, and red chromophore formation in permanently emitting FPs, photoactivatable FPs, and fluorescent timers. The second article focuses on novel monomeric RFPs and their application for studying gene expression, nuclear localization, and dynamics using advanced imaging. https://www.leica-microsystems.com//science-lab/modern-fluorescent-proteins-and-their-biological-applications/ Fri, 18 Nov 2011 12:50:00 +0000 https://www.leica-microsystems.com/3669 Coherent Raman Scattering (CRS) Multiphoton Microscopy Good Vibrations In recent years, new molecular imaging techniques, such as coherent anti-Stokes Raman scattering microscopy (CARS), have been developed for rapid vibrational imaging of living cells. https://www.leica-microsystems.com//science-lab/good-vibrations/ Tue, 12 Apr 2011 22:00:00 +0000 Ph.D. Eric Olaf Potma, Dr. Bernd Sägmüller https://www.leica-microsystems.com/2730 Multiphoton Microscopy Confocal Microscopy Deep Tissue Imaging Developmental biology using Multiphoton microscopy with OPO. To gain new insight into the fundamental control of cell response to physical changes and to study the dynamics and roles of biological flow during the development of the zebrafish, Dr. Julien Vermot established his lab last year at the Institute of Genetics and Molecular and Cellular Biology (IGBMC) in Strasbourg, France. https://www.leica-microsystems.com//science-lab/deep-tissue-imaging/ Mon, 01 Nov 2010 23:00:00 +0000 PhD Julien Vermot, Dr. Andrea Mülter https://www.leica-microsystems.com/2732 Confocal Microscopy Multiphoton Microscopy Live-Cell Imaging Neuroscience Exploring the Concert of Neuronal Activities Brain research using Confocal and Multiphoton Microscopy. Using imaging techniques such as confocal and two-photon microscopy, neuronal dendritic arborization of neurons and their synaptic interconnections can be visualized. https://www.leica-microsystems.com//science-lab/exploring-the-concert-of-neuronal-activities/ Mon, 01 Nov 2010 23:00:00 +0000 Ph.D. Randy Bruno, Ph.D. Myriam Gastard https://www.leica-microsystems.com/2731 Confocal Microscopy Multiphoton Microscopy What is an OPO? Multiphoton microscopy with OPO: imaging with excitation wavelengths up to 1.300 nm. Because light scattering is dependent on the wavelength, better tissue penetration can be achieved by using longer excitation wavelengths. This is where excitation with infrared light, two-photon processes, and the OPO (optical parameter oscillator) can dramatically improve image quality. https://www.leica-microsystems.com//science-lab/what-is-an-opo/ Mon, 01 Nov 2010 23:00:00 +0000 Dr. Bernd Sägmüller, Dr. Andrea Mülter https://www.leica-microsystems.com/2743 Coherent Raman Scattering (CRS) Multiphoton Microscopy Confocal Microscopy CARS and Confocal The most important drawback of single-photon and multiphoton confocal microscopy is the need to label the specimen. CARS (Coherent Anti-Stokes Raman Spectroscopy) addresses this issue because it is non-toxic, non-destructive, and minimally invasive. https://www.leica-microsystems.com//science-lab/cars-and-confocal/ Mon, 01 Nov 2010 23:00:00 +0000 Dr. Stefanie Degenhartt, Ph.D. Vanessa Lurquin https://www.leica-microsystems.com/2561 Multiphoton Microscopy Live-Cell Imaging Confocal Microscopy Stem Cell Biology in Cancer Research The comprehension of stem cell biology and its molecular basis is now acquiring paramount importance in cancer research. The need to look at a single, possibly living, cell makes fluorescence microscopy and confocal microscopy invaluable allies in the study of stem cells. https://www.leica-microsystems.com//science-lab/stem-cell-biology-in-cancer-research/ Thu, 22 Apr 2010 22:00:00 +0000 Laura Furia, Angelo Cicalese, Ilaria Rancati, Isabella Ponzanelli, Paolo Bianchini, Prof. Alberto Diaspro, Mario Faretta https://www.leica-microsystems.com/4290 Multiphoton Microscopy Confocal Microscopy Quantitative Fluorescence Image Restoration and Deconvolution From Molecules to Tissues Cancer research using confocal and multiphoton microscopy. Sequencing of the human genome stimulated a radical change in the approach to biomedical research. The comprehension of the mechanisms regulating life gained a scale-up in throughput to speed up the retrieval of data for a global vision of a system of incomparable complexity. https://www.leica-microsystems.com//science-lab/from-molecules-to-tissues/ Thu, 21 Aug 2008 22:00:00 +0000 Mario Faretta, Prof. Alberto Diaspro, Sara Barozzi, Massimiliano Garrè, Ivan Muradore, Andrea Palamidessi, Dario Parazzoli, Simona Ronzoni, Pietro Transidico, Davide Mazza, Ph.D. Illaria Testa https://www.leica-microsystems.com/4394 Confocal Microscopy Quantitative Fluorescence Multiphoton Microscopy 4D Photoactivation of pa-GFP in Living Cells Using Two-Photon Excitation Laser Scanning Microscopy We report about two-photon activation of a photoactivatable derivative of the Aequorea Victoria green fluorescent protein (pa-GFP). This special form of the molecule increases its fluorescence intensity when excited by 488 nm after irradiation with high intensity light at 413 nm. Two-photon photoactivation produces an effective real three-dimensional (3D) localization of the molecular switching of pa-GFP in the bright state. https://www.leica-microsystems.com//science-lab/4d-photoactivation-of-pa-gfp-in-living-cells-using-two-photon-excitation-laser-scanning-microscopy/ Sat, 14 Oct 2006 22:00:00 +0000 Prof. Alberto Diaspro, Ph.D. Illaria Testa, Valentina Caorsi, Davide Mazza, Ph.D. Giuseppe Vicidomini, Sara Barozzi, Dario Parazzoli, Pietro Transidico, Massimiliano Garrè, Mario Faretta https://www.leica-microsystems.com/6771 Multiphoton Microscopy Confocal Microscopy Multi-photon Excitation Microscopy Advanced microscopical techique for life science: multiphoton microscopy. Multi-photon excitation (MPE) microscopy plays a growing role among microscopical techniques utilized for studying biological matter. In conjunction with confocal microscopy it can be considered the imaging workhorse of life science laboratories. Its roots can be found in a fundamental work written by Maria Goeppert Mayer more than 70 years ago. Nowadays, 2PE and MPE microscopes are expected to increase their impact in areas such biotechnology, neurobiology, embryology, tissue engineering, materials science where imaging can be coupled to the possibility of using the microscopes in an active way, too. https://www.leica-microsystems.com//science-lab/multi-photon-excitation-microscopy/ Mon, 05 Jun 2006 22:00:00 +0000 Prof. Alberto Diaspro, Paolo Bianchini, Ph.D. Giuseppe Vicidomini, Mario Faretta, Paola Ramoino, Cesare Usai