Multidimensional Fluorescence Imaging and Super-Resolution Exploiting Ultrafast Laser and Supercontinuum Technology
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News Release Emmanuelle Charpentier Inducted Into the Hall
News Release Your Contact [email protected] Phone: +49 6151 72-9591 October 22, 2020 Emmanuelle Charpentier Inducted into the Hall of Fame of German Research • Microbiologist, geneticist, infection biologist and Nobel Prize winner Charpentier recognized • Curious Mind Award for young scientists presented to Siegfried Rasthofer and Björn Eskofier Darmstadt, Germany, October 22, 2020– Merck KGaA, Darmstadt, Germany, a leading science and technology company, and manager magazin today inducted Emmanuelle Charpentier (51), Founding Director of the Max Planck Unit for the Science of Pathogens in Berlin, Germany, into the Hall of Fame of German Research. In addition, the two hosts presented the Curious Mind Researcher Award at the same event. Siegfried Rasthofer (32), a computer scientist, received the prize worth € 7,500 in the “Digitalization & Robotics” category. Björn Eskofier (40), an electrical engineer, was also recognized with € 7,500 for his work in the “Life Science” category. In a video message, German Federal Chancellor Angela Merkel said: “It is a privilege to now be able to induct a renowned scientist and designated Nobel laureate into the Hall of Fame of German Research. The Curious Mind Researcher Award also demonstrates that Germany is a research location that offers superb framework conditions for cutting-edge research,” she added and congratulated the prizewinners. “Many scientists are making extraordinary accomplishments – particularly here in Germany as well. This is evident not only in the fight against Covid-19. Thanks to Page 1 of 3 Frankfurter Strasse 250 Head of Media Relations -6328 64293 Darmstadt · Germany Spokesperson: -9591 / -8908 / -45946 / -55707 Hotline +49 6151 72-5000 www.emdgroup.com News Release their passion and perseverance, they are creating the preconditions for the advancement of society. -
Two-Photon Excitation, Fluorescence Microscopy, and Quantitative Measurement of Two-Photon Absorption Cross Sections
Portland State University PDXScholar Dissertations and Theses Dissertations and Theses Fall 12-1-2017 Two-Photon Excitation, Fluorescence Microscopy, and Quantitative Measurement of Two-Photon Absorption Cross Sections Fredrick Michael DeArmond Portland State University Let us know how access to this document benefits ouy . Follow this and additional works at: https://pdxscholar.library.pdx.edu/open_access_etds Part of the Physics Commons Recommended Citation DeArmond, Fredrick Michael, "Two-Photon Excitation, Fluorescence Microscopy, and Quantitative Measurement of Two-Photon Absorption Cross Sections" (2017). Dissertations and Theses. Paper 4036. 10.15760/etd.5920 This Dissertation is brought to you for free and open access. It has been accepted for inclusion in Dissertations and Theses by an authorized administrator of PDXScholar. For more information, please contact [email protected]. Two-Photon Excitation, Fluorescence Microscopy, and Quantitative Measurement of Two-Photon Absorption Cross Sections by Fredrick Michael DeArmond A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Applied Physics Dissertation Committee: Erik J. Sánchez, Chair Erik Bodegom Ralf Widenhorn Robert Strongin Portland State University 2017 ABSTRACT As optical microscopy techniques continue to improve, most notably the development of super-resolution optical microscopy which garnered the Nobel Prize in Chemistry in 2014, renewed emphasis has been placed on the development and use of fluorescence microscopy techniques. Of particular note is a renewed interest in multiphoton excitation due to a number of inherent properties of the technique including simplified optical filtering, increased sample penetration, and inherently confocal operation. With this renewed interest in multiphoton fluorescence microscopy, comes increased interest in and demand for robust non-linear fluorescent markers, and characterization of the associated tool set. -
2–10 Μm Mid-Infrared Fiber-Based Supercontinuum Laser Source
2–10 µm Mid-Infrared Fiber-Based Supercontinuum Laser Source: Experiment and Simulation Sebastien Venck, François St-Hilaire, L Brilland, Amar Nath Ghosh, Radwan Chahal, Céline Caillaud, Marcello Meneghetti, J Troles, Franck Joulain, Solenn Cozic, et al. To cite this version: Sebastien Venck, François St-Hilaire, L Brilland, Amar Nath Ghosh, Radwan Chahal, et al.. 2–10 µm Mid-Infrared Fiber-Based Supercontinuum Laser Source: Experiment and Simulation. Laser & Photonics Reviews, 2020. hal-03023809 HAL Id: hal-03023809 https://hal.archives-ouvertes.fr/hal-03023809 Submitted on 25 Nov 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. 2-10 µm Mid-Infrared All-Fiber Supercontinuum Laser Source: Experiment and Simulation Sébastien Venck1, François St-Hilaire2,6, Laurent Brilland1, Amar N. Ghosh2, Radwan Chahal1, Céline Caillaud1, Marcello Meneghetti3, Johann Troles3, Franck Joulain4, Solenn Cozic4, Samuel Poulain4, Guillaume Huss5, Martin Rochette6, John Dudley2, and Thibaut Sylvestre∗2 1SelenOptics, Campus de Beaulieu, Rennes, France 2Institut FEMTO-ST, -
Second Harmonic Imaging Microscopy
170 Microsc Microanal 9(Suppl 2), 2003 DOI: 10.1017/S143192760344066X Copyright 2003 Microscopy Society of America Second Harmonic Imaging Microscopy Leslie M. Loew,* Andrew C. Millard,* Paul J. Campagnola,* William A. Mohler,* and Aaron Lewis‡ * Center for Biomedical Imaging Technology, University of Connecticut Health Center, Farmington, CT 06030-1507 USA ‡ Division of Applied Physics, Hebrew University of Jerusalem, Jerusalem 91904, Israel Second Harmonic Generation (SHG) has been developed in our laboratories as a high- resolution non-linear optical imaging microscopy (“SHIM”) for cellular membranes and intact tissues. SHG is a non-linear process that produces a frequency doubling of the intense laser field impinging on a material with a high second order susceptibility. It shares many of the advantageous features for microscopy of another more established non-linear optical technique: two-photon excited fluorescence (TPEF). Both are capable of optical sectioning to produce 3D images of thick specimens and both result in less photodamage to living tissue than confocal microscopy. SHG is complementary to TPEF in that it uses a different contrast mechanism and is most easily detected in the transmitted light optical path. It also does not arise via photon emission from molecular excited states, as do both 1- and 2-photon excited fluorescence. SHG of intrinsic highly ordered biological structures such as collagen has been known for some time but only recently has the full potential of high resolution 3D SHIM been demonstrated on live cells and tissues. For example, Figure 1 shows SHIM from microtubules in a living organism, C. elegans. The images were obtained from a transgenic nematode that expresses a ß-tubulin-green fluorescent protein fusion and Figure 1 also shows the TPEF image from this molecule for comparison. -
Supercontinuum Generation in Optical Fibers and Its Biomedical Applications
1/100 Supercontinuum Generation in Optical Fibers and its Biomedical Applications Govind P. Agrawal The Institute of Optics University of Rochester Rochester, New York, USA JJ II J c 2014 G. P. Agrawal I Back Close Introduction • Optical Fibers were developed during the 1960s with medical appli- cations in mind (endoscopes). 2/100 • During 1980{2000 optical fibers were exploited for telecommunica- tions and now form the backbone for the Internet. • Biomedical applications of fibers increased after 2000 with the ad- vent of photonic crystal and other microstructured fibers. • Supercontinuum (ultrabroad coherent spectrum) is critical for many biomedical applications. • Nonlinear effects inside fibers play an important role in generating JJ a supercontinuum. II • This talk focuses on Supercontinuum generation with emphasis on J I their biomedical applications. Back Close Supercontinuum History • Discovered in 1969 using borosilicate glass as a nonlinear medium 3/100 [Alfano and Shapiro, PRL 24, 584 (1970)]. • In this experiment, 300-nm-wide supercontinuum covered the entire visible region. • A 20-m-long fiber was employed in 1975 to produce 180-nm wide supercontinuum using Q-switched pulses from a dye laser [Lin and Stolen, APL 28, 216 (1976)]. • 25-ps pulses were used in 1987 but the bandwidth was only 50 nm [Beaud et al., JQE 23, 1938 (1987)]. JJ • 200-nm-wide supercontinuum obtained in 1989 by launching 830-fs II pulses into 1-km-long single-mode fiber [Islam et al., JOSA B 6, J 1149 (1989)]. I Back Close Supercontinuum History • Supercontinuum work with optical fibers continued during 1990s 4/100 with telecom applications in mind. -
Fv1000 Fluoview
Confocal Laser Scanning Biological Microscope FV1000 FLUOVIEW FLUOVIEW—Always Evolving FLUOVIEW–—From Olympus is Open FLUOVIEW—More Advanced than Ever The Olympus FLUOVIEW FV1000 confocal laser scanning microscope delivers efficient and reliable performance together with the high resolution required for multi-dimensional observation of cell and tissue morphology, and precise molecular localization. The FV1000 incorporates the industry’s first dedicated laser light stimulation scanner to achieve simultaneous targeted laser stimulation and imaging for real-time visualization of rapid cell responses. The FV1000 also measures diffusion coefficients of intracellular molecules, quantifying molecular kinetics. Quite simply, the FLUOVIEW FV1000 represents a new plateau, bringing “imaging to analysis.” Olympus continues to drive forward the development of FLUOVIEW microscopes, using input from researchers to meet their evolving demands and bringing “imaging to analysis.” Quality Performance with Innovative Design FV10i 1 Imaging to Analysis ing up New Worlds From Imaging to Analysis FV1000 Advanced Deeper Imaging with High Resolution FV1000MPE 2 Advanced FLUOVIEW Systems Enhance the Power of Your Research Superb Optical Systems Set the Standard for Accuracy and Sensitivity. Two types of detectors deliver enhanced accuracy and sensitivity, and are paired with a new objective with low chromatic aberration, to deliver even better precision for colocalization analysis. These optical advances boost the overall system capabilities and raise performance to a new level. Imaging, Stimulation and Measurement— Advanced Analytical Methods for Quantification. Now equipped to measure the diffusion coefficients of intracellular molecules, for quantification of the dynamic interactions of molecules inside live cell. FLUOVIEW opens up new worlds of measurement. Evolving Systems Meet the Demands of Your Application. -
Towards On-Chip Self-Referenced Frequency-Comb Sources Based on Semiconductor Mode-Locked Lasers
micromachines Review Towards On-Chip Self-Referenced Frequency-Comb Sources Based on Semiconductor Mode-Locked Lasers Marcin Malinowski 1,*, Ricardo Bustos-Ramirez 1 , Jean-Etienne Tremblay 2, Guillermo F. Camacho-Gonzalez 1, Ming C. Wu 2 , Peter J. Delfyett 1,3 and Sasan Fathpour 1,3,* 1 CREOL, The College of Optics and Photonics, University of Central Florida, Orlando, FL 32816, USA; [email protected] (R.B.-R.); [email protected] (G.F.C.-G.); [email protected] (P.J.D.) 2 Department of Electrical Engineering and Computer Sciences, University of California, Berkeley, CA 94720, USA; [email protected] (J.-E.T.); [email protected] (M.C.W.) 3 Department of Electrical and Computer Engineering, University of Central Florida, Orlando, FL 32816, USA * Correspondence: [email protected] (M.M.); [email protected] (S.F.) Received: 14 May 2019; Accepted: 5 June 2019; Published: 11 June 2019 Abstract: Miniaturization of frequency-comb sources could open a host of potential applications in spectroscopy, biomedical monitoring, astronomy, microwave signal generation, and distribution of precise time or frequency across networks. This review article places emphasis on an architecture with a semiconductor mode-locked laser at the heart of the system and subsequent supercontinuum generation and carrier-envelope offset detection and stabilization in nonlinear integrated optics. Keywords: frequency combs; heterogeneous integration; second-harmonic generation; supercontinuum; integrated photonics; silicon photonics; mode-locked lasers; nonlinear optics 1. Introduction The field of integrated photonics aims at harnessing optical waves in submicron-scale devices and circuits, for applications such as transmitting information (communications) and gathering information about the environment (imaging, spectroscopy, etc.). -
Second Harmonic Generation Microscopy for Quantitative Analysis
PROTOCOL Second harmonic generation microscopy for quantitative analysis of collagen fibrillar structure Xiyi Chen1, Oleg Nadiarynkh1,2, Sergey Plotnikov1,2 & Paul J Campagnola1 1Department of Biomedica l Engineering, University of Wisconsin-Madison, Madison, Wisconsin, USA. 2Present addresses: Department of Physics, University of Utrecht, Utrecht, The Netherlands (O.N.); US National Institutes of Health, Heart, Lung and Blood Institute, Bethesda, Maryland, USA (S.P.). Correspondence should be addressed to P.J.C. ([email protected]). Published online 8 March 2012; doi:10.1038/nprot.2012.009 Second-harmonic generation (SHG) microscopy has emerged as a powerful modality for imaging fibrillar collagen in a diverse range of tissues. Because of its underlying physical origin, it is highly sensitive to the collagen fibril/fiber structure, and, importantly, to changes that occur in diseases such as cancer, fibrosis and connective tissue disorders. We discuss how SHG can be used to obtain more structural information on the assembly of collagen in tissues than is possible by other microscopy techniques. We first provide an overview of the state of the art and the physical background of SHG microscopy, and then describe the optical modifications that need to be made to a laser-scanning microscope to enable the measurements. Crucial aspects for biomedical applications are the capabilities and limitations of the different experimental configurations. We estimate that the setup and calibration of the SHG instrument from its component parts will require 2–4 weeks, depending on the level of the user′s experience. INTRODUCTION Over the past decade, the nonlinear optical method of SHG micros on the harmonophores and their assembly that can be imaged, as copy has emerged as a powerful tool for visualizing the supra the environment must be noncentrosymmetric on the size scale molecular assembly of collagen in tissues at an unprecedented level of of λSHG; otherwise, the signal will vanish. -
STED Fluorescence Microscopy: a Method of Resolution Enhancement Submitted by David Biss and Jason Neiser
STED Fluorescence Microscopy: A method of resolution enhancement Submitted by David Biss and Jason Neiser Introduction relaxed vibrational level of the ground electronic state. The microscope excitation light generates If geometrical aberrations are minimized in an a transition in the fluorophore from level L0 to optical system, the smallest spot size attainable is L1, a high vibrational level of the first excited the diffraction limited spot size. Confocal state. From here, the molecule undergoes a fast microscopy was the first method to extend vibrational decay from L1 to L2, and eventually resolution beyond the Abbe resolution limit and fluoresces to L3 by spontaneous emission. it added axial resolution to the system. This form of microscopy images a portion of the sample being investigated onto a confocal pinhole at the detection plane of the system. Since the invention of confocal microscopy other methods have been devised to reach beyond the standard diffraction limit. Some of these methods are 4π microscopy, two photon microscopy, near-field microscopy, and more recently, STimulated Emission Depletion (STED) fluorescence microscopy. [1, 2, 3] STED fluorescence microscopy takes standard fluorescence microscopy and introduces a technique to reduce the emitted spot size. STED microscopy uses stimulated emission to deplete fluorophores before they fluoresce. If this depletion occurs at the edges of the excited sample area the spot size (and volume) of the fluorescence can be reduced beyond the Fig. 1 Energy level diagram of a dye molecule. A short diffraction limit. wavelength pulse excites the molecule and it may be relaxed by either fluorescing or by stimulated emission via the STED Theory pulse. -
Practical Tips for Two-Photon Microscopy
Appendix 1 Practical Tips for Two-Photon Microscopy Mark B. Cannell, Angus McMorland, and Christian Soeller INTRODUCTION blue and green diode lasers. To provide an alignment beam to which the external laser can be aligned, light from this reference As is clear from a number of the chapters in this volume, 2-photon laser needs to be bounced back through the microscope optical microscopy offers many advantages, especially for living-cell train and out through the external coupling port: studies of thick specimens such as brain slices and embryos. CAUTION: Before you switch on the reference laser in this However, these advantages must be balanced against the fact that configuration make sure that all PMTs are protected and/or commercial multiphoton instrumentation is much more costly than turned off. the equipment used for confocal or widefield/deconvolution. Given Place a front-surface mirror on the stage of the microscope and these two facts, it is not surprising that, to an extent much greater focus onto the reflective surface using an air objective for conve- than is true of confocal, many researchers have decided to add a nience (at sharp focus, you should be able to see scratches or other femtosecond (fs) pulsed near-IR laser to a scanner and a micro- mirror defects through the eyepieces). The idea of this method is scope to make their own system (Soeller and Cannell, 1996; Tsai to cause the reference laser beam to bounce back through the et al., 2002; Potter, 2005). Even those who purchase a commercial optical train and emerge from the other laser port. -
State-Of-The-Art Fiber Optics for Short Distance Frequency Reference Distribution
N89-27878 January-March 1989 TDA Progress Report 42-97 State-of-the-Art Fiber Optics for Short Distance Frequency Reference Distribution G. Lutes and L. Primas Communications Systems Research Section A number of recently developed fiber-optic components that hem the promise of unprecedented stability for passively stabilized frequency distribution links are character- ized. These components include a fiber-optic transmitter, an optical isolator, and a new type of fiber-optic cable. A novel laser transmitter exhibits extremely low sensitivity to intensity and polarization changes of reflected light due to cable flexure. This virtually eliminates one of the shortcomings in previous laser transmitters. A high-isolation, low- loss optical isolator has been developed which also virtually eliminates laser sensitivity to changes in intensity and polarization of reflected light. A newly developed fiber has been tested. This fiber has a thermal coefficient of delay of less than 0.5 parts per million per °C, nearly 20 times lower than the best coaxial hardline cable and 10 times lower than any previous fiber-optic cable. These components are highly suitable for distribution systems with short extent, such as within a Deep Space Communications Complex. In this article these new components are described and the test results presented. I. Introduction mary causes of degradation. These effects are caused by distri- bution system noise, which reduces the SNR, and variations in The transmitter exciter, local oscillator, and receiver delay the environmental temperature, which cause delay changes. calibration system in a Deep Space Station (DSS) require The degree of delay change is dependent on the Thermal Coef- stable frequency references. -
Arrangement of a 4Pi Microscope for Reducing the Confocal Detection
Arrangement of a 4Pi microscope for reducing the confocal detection volume with two-photon excitation Nicolas Sandeau∗and Hugues Giovannini† Received 16 November 2005; received in revised form 7 February 2006; accepted 8 February 2006 Institut Fresnel, UMR 6133 CNRS, Universit´ePaul C´ezanne Aix-Marseille III, F-13397 Marseille cedex 20, France The main advantage of two-photon fluorescence confocal microscopy is the low absorption obtained with live tissues at the wavelengths of operation. However, the resolution of two-photon fluorescence confocal microscopes is lower than in the case of one-photon excitation. The 4Pi microscope type C working in two-photon regime, in which the excitation beams are coherently superimposed and, simultaneously, the emitted beams are also coherently added, has shown to be a good solution for increasing the resolution along the optical axis and for reducing the amplitude of the side lobes of the point spread function. However, the resolution in the transverse plane is poorer than in the case of one-photon excitation due to the larger wavelength in- volved in the two-photon fluorescence process. In this paper we show that a particular arrangement of the 4Pi microscope, referenced as 4Pi’ microscope, is a solution for obtaining a lateral resolution in the two-photon regime sim- ilar or even better to that obtained with 4Pi microscopes working in the arXiv:physics/0610036v1 [physics.bio-ph] 6 Oct 2006 one-photon excitation regime. Keywords: Resolution; Fluorescence microscopy; 4Pi microscopy; Confocal; Detection volume; Two-photon excitation ∗E-mail: [email protected] †E-mail: [email protected]; Tel: +33 491 28 80 66; Fax: +33 491 28 80 67 1 1 Introduction Strong efforts have been made in the last decade to improve the resolution of fluorescence microscopes.