Leica Science Lab - Tag : Multiphoton Imaging https://www.leica-microsystems.com//science-lab/tag/tags/multiphoton-imaging/show/Tag/ Article tagged with Multiphoton Imaging en-US https://www.leica-microsystems.com/4421 Coherent Raman Scattering (CRS) Coherent Raman Scattering Microscopy Publication List CRS (Coherent Raman Scattering) microscopy is an umbrella term for label-free methods that image biological structures by exploiting the characteristic, intrinsic vibrational contrast of their molecules. The two most important CRS techniques are Coherent Anti-Stokes Raman Scattering (CARS) and Stimulated Raman Scattering (SRS). The biochemical image contrast of CRS is in many ways complementary to the molecular contrast obtained in fluorescence microscopy. A second crucial advantage of these methods is that they preserve the specimen/sample in a near pristine state. This reference list presents current and basic papers on CRS microscopy. https://www.leica-microsystems.com//science-lab/cars-publication-list/ Tue, 08 Oct 2019 22:00:00 +0000 Dr. Volker Schweikhard https://www.leica-microsystems.com/20418 Coherent Raman Scattering (CRS) Multiphoton Microscopy Direct visualization of the arterial wall water permeability barrier using CARS microscopy The artery wall is equipped with a water permeation barrier that allows blood to flow at high pressure without significant water leak. The precise location of this barrier is unknown despite its importance in vascular function and its contribution to many vascular complications when it is compromised. Herein we map the water permeability in intact arteries, using coherent anti-Stokes Raman scattering (CARS) microscopy and isotopic perfusion experiments. Generation of the CARS signal is optimized for water imaging with broadband excitation. https://www.leica-microsystems.com//science-lab/direct-visualization-of-the-arterial-wall-water-permeability-barrier-using-cars-microscopy/ Wed, 22 Aug 2018 22:00:00 +0000 https://www.leica-microsystems.com/20323 Confocal Microscopy Which Sensor is the Best for Confocal Imaging? The Hybrid Photodetectors (HyD) are! Why that is the case is explained in this short Science Lab article. https://www.leica-microsystems.com//science-lab/which-sensor-is-the-best-for-confocal-imaging/ Mon, 16 Jul 2018 22:00:00 +0000 Dr. Rolf T. Borlinghaus, Holger Birk 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/19451 Live-Cell Imaging Fluorescence Microscopy Multiphoton Microscopy Five Questions Asked: Prof. Dr. Jacco van Rheenen speaks about the most important considerations when imaging deep into mouse tissue When operating a confocal microscope, or when discussing features and parameters of such a device, we inescapably mention the pinhole and its diameter. This short introductory document is meant to explain the significance of the pinhole for those, who did not want to spend too much time to dig into theory and details of confocal microscopy but wanted to have an idea about the effect of the pinhole. https://www.leica-microsystems.com//science-lab/five-questions-asked-prof-dr-jacco-van-rheenen-speaks-about-the-most-important-considerations-when-imaging-deep-into-mouse-tissue/ Fri, 19 May 2017 05:47:00 +0000 PhD Jacco van Rheenen, Dr. Bernd Sägmüller 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/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/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/10286 Live-Cell Imaging Multiphoton Microscopy Direct In Vivo Evidence for Tumor Propagation by Glioblastoma Cancer Stem Cells Cancer research using multiphoton microscopy. High-grade gliomas (World Health Organization grade III anaplastic astrocytoma and grade IV glioblastoma multiforme), the most prevalent primary malignant brain tumors, display a cellular hierarchy with self-renewing, tumorigenic cancer stem cells (CSCs) at the apex. While the CSC hypothesis has been an attractive model to describe many aspects of tumor behavior, it remains controversial due to unresolved issues including the use of ex vivo analyses with differential growth conditions. https://www.leica-microsystems.com//science-lab/direct-in-vivo-evidence-for-tumor-propagation-by-glioblastoma-cancer-stem-cells/ Fri, 12 Jul 2013 13:21:00 +0000 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/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 Bianco, Prof. Alberto Diaspro, Anna Tampieri, Ugo Valbusa, Giuseppe Firpo, Roberto Marotta, Monica Sandri, Paolo Bianchini, Rodolfo Quarto 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/10190 Quantitative Fluorescence Multiphoton Microscopy A New FRAP/FRAPa Method for Three-Dimensional Diffusion Measurements Based on Multiphoton Excitation Microscopy Quantitative measurement method based on FRAP and FRAPa using multiphoton microscopy. We present a new convenient method for quantitative three-dimensionally resolved diffusion measurements based on the photobleaching (FRAP) or photoactivation (FRAPa) of a disk-shaped area by the scanning laser beam of a multiphoton microscope. Contrary to previously reported spot-photobleaching protocols, this method has the advantage of full scalability of the size of the photobleached area and thus the range of diffusion coefficients, which can be measured conveniently. https://www.leica-microsystems.com//science-lab/a-new-frapfrapa-method-for-three-dimensional-diffusion-measurements-based-on-multiphoton-excitation-microscopy/ Fri, 11 Jul 2008 15:06:00 +0000 Davide Mazza 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, Massimiliano Garrè, Pietro Transidico, Dario Parazzoli, Sara Barozzi, Ph.D. Giuseppe Vicidomini, Davide Mazza, Valentina Caorsi, Ph.D. Illaria Testa, 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