Possibility of Using the Beet Dyes As a Laser Gain Medium
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Absolute Fluorescence Quantum Yields Using the FL 6500 Fluorescence Spectrometer
APPLICATION NOTE Fluorescence Spectroscopy Authors: Kathryn Lawson-Wood Steve Upstone Kieran Evans PerkinElmer, Inc. Seer Green, UK Absolute Fluorescence Quantum Yields Using the FL 6500 Introduction Fluorescence quantum yield Fluorescence Spectrometer (ΦF) is a characteristic property of a fluorescent species and is denoted as the ratio of the number of photons emitted through fluorescence, to the number of photons absorbed by the fluorophore (Equation 1). ΦF ultimately relates to the efficiency of the pathways leading to emission of fluorescence (Figure 1), providing the probability of the excited state being deactivated by fluorescence rather than by other competing relaxation processes. The magnitude of ΦF is directly related to the intensity of the observed fluorescence.1 Number of photons emitted through fluorescence ΦF = (1) Number of photons absorbed Fluorescence quantum yield can be measured using two methods: the relative method and the absolute method. In comparison with relative quantum yield measurements, the absolute method is applicable to both solid and solution phase measurements. The ΦF can be determined in a single measurement without the need for a reference standard. It is especially useful for samples which absorb and emit in wavelength regions for which there are no reliable quantum yield standards available.2 The importance of ΦF has been shown in several industries, standard. It is especially useful for samples which absorb and including the research, development and evaluation of audio emit in wavelength regions for which there are no reliable visual equipment, electroluminescent materials (OLED/LED) and quantum yield standards available. This method requires an fluorescent probes for biological assays.3 This application integrating sphere which allows the instrument to collect all of demonstrates the use of the PerkinElmer FL 6500 Fluorescence the photons emitted from the sample. -
Studies on Fluorescence Efficiency and Photodegradation Of
Laser Chemistry, 2002, Vol. 20(2–4), pp. 99–110 Studies on Fluorescence Efficiency and Photodegradation of Rhodamine 6G Doped PMMA Using a Dual Beam Thermal Lens Technique ACHAMMA KURIAN*, NIBU A GEORGE, BINOY PAUL, V. P. N. NAMPOORI and C. P. G. VALLABHAN International School of Photonics, Cochin University of Science & Technology, Cochin 682022, India (Received11September 2002) In this paper we report the use of the dual beam thermal lens technique as a quantitative method to determine absolute fluorescence quantum efficiency and concentration quenching of fluorescence emission from rhodamine 6G doped Poly(methyl methacrylate) (PMMA), prepared with different concentrations of the dye. A comparison of the present data with that reported in the literature indicates that the observed variation of fluorescence quantum yield with respect to the dye concentration follows a similar profile as in the earlier reported observations on rhodamine 6G in solution. The photodegradation of the dye molecules under cw laser excitation is also studied using the present method. Key words: Thermal lens; Fluorescence quantum yield; Photostability; Dye doped polymer 1 INTRODUCTION From the mid 60s, dye lasers have been attractive sources of coherent tunable radiation because of their unique operational flexibility. The salient features of dye lasers are their tunability, with emission from near ultraviolet to the near infrared. High gain, broad spectral bandwidth enabled pulsed and continuous wave operation. Although dyes have been demonstrated to lase in the solid, liquid or gas phases, liquid solutions of dyes in suitable organic * Corresponding author. Catholicate College, Pathanamthitta 689645, India. E-mail: [email protected] 100 A. KURIAN et al. -
Measurement of Fluorescence Quantum Yields on ISS Instrumentation Using Vinci Yevgen Povrozin and Ewald Terpetschnig
TECHNICAL NOTE Measurement of Fluorescence Quantum Yields on ISS Instrumentation Using Vinci Yevgen Povrozin and Ewald Terpetschnig Introduction The quantum yield Q can also be described by the relative rates of the radiative and non-radiative pathways, which deactivate the excited state: kr Q [1 ] kkr nr where and correspond to radiative and non-radiative processes, respectively. In this equation, kr knr knr describes the sum of the rate constants for the various processes that compete with the emission process. These processes include photochemical, and dissociative processes including other, less well characterized changes that result in a return to the ground state with simultaneous dissipation of the energy of the excited state into heat. These latter processes are collectively called non-radiative transitions and two types have been clearly recognized: intersystem crossing and internal conversion. Intersystem crossing related to the radiationless spin inversion of a singlet state (S1) in the excited state into a triplet state (T1). The luminescence quantum yield is also given as the ratio of the number of photons emitted to the number of photons absorbed by the sample: photonsem Q [ 2] photonsabs While measurements of the “absolute” quantum yield do require more sophisticated instrumentation [1], it is easier to determine the “relative” quantum yield of a fluorophore by comparing it to a standard with a known quantum yield. Some of the most common standards are listed in table below: Conditions for Quantum Yield [Q.Y.] Q.Y. Excitation Q.Y. Ref. Standards [nm] [%] Measurements Cy3 4 PBS 540 2 Cy5 27 PBS 620 2 Cresyl Violet 53 Methanol 580 3 1 ISS TECHNICAL NOTE Conditions for Quantum Yield [Q.Y.] Q.Y. -
Increased Mitochondrial Uptake of Rhodamine 123 During Lymphocyte Stimulation
Proc. Natl. Acad. Sci. USA Vol. 78, No. 4, pp. 2383-2387, April 1981 Cell Biology Increased mitochondrial uptake of rhodamine 123 during lymphocyte stimulation (flow cytometry/cycling-noncycling cells/supravital fluorescent probe) ZBIGNIEW DARZYNKIEWICZ, LISA STAIANO-COICO, AND MYRON R. MELAMED Memorial Sloan-Kettering Cancer Center, New York, New York 10021 Communicated by Lloyd J. Old, January 5, 1981 ABSTRACT The positively charged rhodamine analog rho- trifugation on Ficoll/Isopaque (Lymphoprep; Nyegaardo, Oslo, damine 123 accumulates specifically in the mitochondria of living Norway). The mononuclear cells were suspended in Eagle's cells. In the present work, the uptake of rhodamine 123 by indi- basal medium containing 15% fetal bovine serum and subcul- vidual lymphocytes undergoing blastogenic transformation in cul- tured in plastic dishes to remove most ofthe monocytes (3). The tures stimulated by phytohemagglutinin was measured by flow nonadhering cells were then suspended in Eagle's medium con- cytometry. A severalfold increase in cell ability to accumulate rho- taining penicillin, streptomycin, and 15% fetal bovine serum damine 123 was observed during lymphocyte stimulation. Maxi- and incubated at 37°C in the presence of PHA (phytohemag- mal dye uptake, seen on the third day ofcell stimulation, coincided glutinin M, GIBCO) as described (2, 3). The samples were with- in time with the peak of DNA synthesis (maximal number of cells drawn from these cultures at daily intervals. Other details ofthe in the S phase) and mitotic activity. A large intercellular variation are elsewhere 3). among stimulated lymphocytes, with some cells having fluores- cell culture system given (2, cence increased as much as 15 times in comparison with nonstim- Cell Staining. -
Reversal of Resistance to Rhodamine 123 in Adriamycin-Resistant Friend Leukemia Cells1
[CANCER RESEARCH 45, 2626-2631, June 1985] Reversal of Resistance to Rhodamine 123 in Adriamycin-resistant Friend Leukemia Cells1 Theodore J. Lampidis,2 Jean-Nicolas Munck, Awtar Krishan, and Haim Tapiero Department of Oncology, University of Miami, School of Medicine, Comprehensive Cancer Center for the State of Florida, Miami, Florida 33101 [T. J. L, A. K.], and Departement de Pharmacologie Cellulaire, Moléculaireet de Pharmacocinétique, ICIG (CNRS LA-149), Hôpital Paul-Brousse, 94804 Villejuif, France [J-N. M., H. T.] ABSTRACT the calcium transport inhibitor, verapamil, on the cellular accu mulation and retention of Rho-123 and ADM as it relates to Pleiotropic resistance to rhodamine 123 (Rho-123) in Adria- resistance in these cell variants. mycin (ADM)-resistant Friend leukemia cells was circumvented by cotreatment with 10 //M verapamil. Increased cytotoxicity corresponded to higher intracellular Rho-123 levels. The vera- MATERIALS AND METHODS pamil-induced increase of drug accumulation in resistant cells is Cells and Cytotoxicity Assay. FLC were derived from a clone of accounted for at least in part by the blockage or slowing of Rho- Friend virus-tranformed 745A cells. Cells were seeded and grown as 123 efflux from these cells. In contrast, accumulation and con described previously (8). The cell variant resistant to ADM (ADM-RFLC3) sequent cytotoxicity of Rho-123 in sensitive cells are not in was derived from the above clone by continuous exposure to ADM (8). creased by verapamil. Similar results were obtained when ADM Resistant cells, grown for more than 150 passages in drug-free medium, was used in this cell system. -
Photoremovable Protecting Groups Used for the Caging of Biomolecules
1 1 Photoremovable Protecting Groups Used for the Caging of Biomolecules 1.1 2-Nitrobenzyl and 7-Nitroindoline Derivatives John E.T. Corrie 1.1.1 Introduction 1.1.1.1 Preamble and Scope of the Review This chapter covers developments with 2-nitrobenzyl (and substituted variants) and 7-nitroindoline caging groups over the decade from 1993, when the author last reviewed the topic [1]. Other reviews covered parts of the field at a similar date [2, 3], and more recent coverage is also available [4–6]. This chapter is not an exhaustive review of every instance of the subject cages, and its principal focus is on the chemistry of synthesis and photocleavage. Applications of indi- vidual compounds are only briefly discussed, usually when needed to put the work into context. The balance between the two cage types is heavily slanted to- ward the 2-nitrobenzyls, since work with 7-nitroindoline cages dates essentially from 1999 (see Section 1.1.3.2), while the 2-nitrobenzyl type has been in use for 25 years, from the introduction of caged ATP 1 (Scheme 1.1.1) by Kaplan and co-workers [7] in 1978. Scheme 1.1.1 Overall photolysis reaction of NPE-caged ATP 1. Dynamic Studies in Biology. Edited by M. Goeldner, R. Givens Copyright © 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim ISBN: 3-527-30783-4 2 1 Photoremovable Protecting Groups Used for the Caging of Biomolecules 1.1.1.2 Historical Perspective The pioneering work of Kaplan et al. [7], although preceded by other examples of 2-nitrobenzyl photolysis in synthetic organic chemistry, was the first to apply this to a biological problem, the erythrocytic Na:K ion pump. -
(12) United States Patent (10) Patent No.: US 7476,504 B2 Turner (45) Date of Patent: Jan
USOO7476504B2 (12) United States Patent (10) Patent No.: US 7476,504 B2 Turner (45) Date of Patent: Jan. 13, 2009 (54) USE OF REVERSIBLE EXTENSION Glatthar, Ralf, et al. 2000. A New Photocleavable Linker in Solid TERMINATOR IN NUCLECACD Phase Chemistry for Ether Cleavage. Org. Lett. 2(15): 2315-2317. SEQUENCING Goldmacher, Victor S., et al. 1992. Photoactivation of Toxin Conju gates. Bioconjugate Chem. 3(2): 104-107. Guillier, Fabrice, et al. 2000. Linkers and Cleavage Strategies in (75) Inventor: Stephen Turner, Menlo Park, CA (US) Solid-Phase Organic Synthesis and Combinatorial Chemistry. Chem. Rev. 100(6): 2091-2157. (73) Assignee: Pacific Biosciences of California, Inc., Holmes, Christopher P. 1997. Model Studies for New o-Nitrobenzyl Menlo Park, CA (US) Photolabile Linkers: Substituent Effects on the Rates of Photochemi cal Cleavage.J. Org. Chem. 62(8): 2370-2380. (*) Notice: Subject to any disclaimer, the term of this Hu, Jun, et al. 2001. Competitive Photochemical Reactivity in a patent is extended or adjusted under 35 Self-Assembled Monolayer on a Colloidal Gold Cluster. J. Am. U.S.C. 154(b) by 365 days. Chem. Soc. 123(7): 1464-1470. Kessler, Martin, et al. 2003. Sequentially Photocleavable Protecting (21) Appl. No.: 11/341,041 Groups in Solid-Phase Synthesis. Org. Lett.5(8); 1179-1181. Kim, Hanyoung, et al. 2004. Core-Shell-Type Resins for Solid-Phase (22) Filed: Jan. 27, 2006 Peptide Synthesis: Comparison with Gel-Type Resins in Solid-Phase Photolytic Cleavage Reaction. Org. Lett. 6(19): 3273-3276. (65) Prior Publication Data Maxam, Allan M., et al. 1977. A new method for sequencing DNA. -
Two-Photon Excitation Fluorescence Microscopy
P1: FhN/ftt P2: FhN July 10, 2000 11:18 Annual Reviews AR106-15 Annu. Rev. Biomed. Eng. 2000. 02:399–429 Copyright c 2000 by Annual Reviews. All rights reserved TWO-PHOTON EXCITATION FLUORESCENCE MICROSCOPY PeterT.C.So1,ChenY.Dong1, Barry R. Masters2, and Keith M. Berland3 1Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139; e-mail: [email protected] 2Department of Ophthalmology, University of Bern, Bern, Switzerland 3Department of Physics, Emory University, Atlanta, Georgia 30322 Key Words multiphoton, fluorescence spectroscopy, single molecule, functional imaging, tissue imaging ■ Abstract Two-photon fluorescence microscopy is one of the most important re- cent inventions in biological imaging. This technology enables noninvasive study of biological specimens in three dimensions with submicrometer resolution. Two-photon excitation of fluorophores results from the simultaneous absorption of two photons. This excitation process has a number of unique advantages, such as reduced specimen photodamage and enhanced penetration depth. It also produces higher-contrast im- ages and is a novel method to trigger localized photochemical reactions. Two-photon microscopy continues to find an increasing number of applications in biology and medicine. CONTENTS INTRODUCTION ................................................ 400 HISTORICAL REVIEW OF TWO-PHOTON MICROSCOPY TECHNOLOGY ...401 BASIC PRINCIPLES OF TWO-PHOTON MICROSCOPY ..................402 Physical Basis for Two-Photon Excitation ............................ -
Photochemistry
Photochemistry 1.1 Introduction:- Photochemistry is concerned with reactions which are initiated by electronically excited molecules. Such molecules are produced by the absorption of suitable radiation in the visible and near ultraviolet region of the spectrum. Photochemistry is basic to the world we live in with sun as the central figure, the origin of life must have been a photochemical act. Simple gaseous molecules like methane, ammonia and carbon dioxide must have reacted photochemically to synthesize complex organic molecules like proteins and nucleic acids. Photobiology, the photochemistry of biological reactions, is a rapidly developing subject and helps in understanding the phenomenon of photosynthesis, phototaxin, photoperiodism, vision and mutagenic effects of light. The relevance of photochemistry also lies in its varied applications in science and technology. Synthetic organic photochemistry has provided methods for the manufacture of many chemicals which could not be produced by dark reactions. Some industrially viable photochemical syntheses include synthesis of vitamin D2 from ergosterol isolated from yeast, synthesis of caprolactum which is the monomer for Nylon 6, manufacture of cleaning solvents and synthesis of some antioxidants. Photoinitiated polymerization and photopolymerisation are used in photography, lithoprinting and manufacture of printed circuits for the electronic industry. The photophysical phenomena of flourescence and phosphorescence have found varied applications in fluorescent tube lights, TV screens, as luminescent dials for watches, as “optical brighteners” in white dress materials, as paints in advertisement hoardings and so on. Another revolutionary application of electronically excited molecular systems is laser technology. The two main processes, therefore, studied under photochemistry are: 1. Photophysical process 2. Photochemical process 1. -
Lifetime and Fluorescence Quantum Yield of Two Fluorescein-Amino Acid-Based Compounds in Different Organic Solvents and Gold Colloidal Suspensions
chemosensors Article Lifetime and Fluorescence Quantum Yield of Two Fluorescein-Amino Acid-Based Compounds in Different Organic Solvents and Gold Colloidal Suspensions Viviane Pilla 1,2,*, Augusto C. Gonçalves 1,3, Alcindo A. Dos Santos 3 and Carlos Lodeiro 1,4 ID 1 BIOSCOPE Group, LAQV-REQUIMTE, Chemistry Department, Faculty of Science and Technology, University NOVA of Lisbon, 2829-516 Caparica, Portugal; [email protected] (A.C.G.); [email protected] (C.L.) 2 Instituto de Física, Universidade Federal de Uberlândia-UFU, Av. João Naves de Ávila 2121, 38400-902 Uberlândia, Brazil 3 Instituto de Química, Universidade de São Paulo-USP, Av. Prof. Lineu Prestes 748, CxP 26077, 05508-000 São Paulo, Brazil; [email protected] 4 Proteomass Scientific Society, Rua dos Inventores, Madan Park, 2829-516 Caparica, Portugal * Correspondence: [email protected] Received: 11 May 2018; Accepted: 28 June 2018; Published: 30 June 2018 Abstract: Fluorescein and its derivatives are yellowish-green emitting dyes with outstanding potential in advanced molecular probes for different applications. In this work, two fluorescent compounds containing fluorescein and an amino acid residue (compounds 1 and 2) were photophysically explored. Compounds 1 and 2 were previously synthesized via ester linkage between fluorescein ethyl ester and Boc-ser (TMS)-OH or Boc-cys(4-methyl benzyl)-OH. Studies on the time-resolved fluorescence lifetime and relative fluorescence quantum yield (φ) were performed for 1 and 2 diluted in acetonitrile, ethanol, dimethyl sulfoxide, and tetrahydrofuran solvents. The discussion considered the dielectric constants of the studied solvents (range between 7.5 and 47.2) and the photophysical parameters. -
Mediated Multidrug Resistance (MDR) in Cancer
A Thesis Entitled: Differential Effects of Dopamine D3 Receptor Antagonists in Modulating ABCG2 - Mediated Multidrug Resistance (MDR) in Cancer by Noor A. Hussein Submitted to the Graduate Faculty as partial fulfillment of the requirements for the Master of Science in Pharmaceutical Sciences Degree in Pharmacology and Toxicology _________________________________________ Dr. Amit K. Tiwari, Committee Chair _________________________________________ Dr. Frank Hall , Committee Member _________________________________________ Dr. Zahoor Shah, Committee Member _________________________________________ Dr. Caren Steinmiller, Committee Member _________________________________________ Dr. Amanda Bryant-Friedrich, Dean College of Graduate Studies The University of Toledo May 2017 Copyright 2017, Noor A. Hussein This document is copyrighted material. Under copyright law, no parts of this document may be reproduced without the expressed permission of the author. An Abstract of Differential Effects of Dopamine D3 Receptor Antagonists in Modulating ABCG2 - Mediated Multidrug Resistance (MDR) by Noor A. Hussein Submitted to the Graduate Faculty as partial fulfillment of the requirements for the Master of Science in Pharmaceutical Sciences Degree in Pharmacology and Toxicology The University of Toledo May 2017 The G2 subfamily of the ATP-binding cassette transporters (ABCG2), also known as the breast cancer resistance protein (BRCP), is an efflux transporter that plays an important role in protecting the cells against endogenous and exogenous toxic substances. The ABCG2 transporters are also highly expressed in the blood brain barrier (BBB), providing protection against specific toxic compounds. Unfortunately, their overexpression in cancer cells results in the development of multi-drug resistance (MDR), and thus, chemotherapy failure. Dopamine3 receptor (D3R) antagonists were shown to have excellent anti-addiction properties in preclinical animal models but produced limited clinical success with the lead molecule. -
Selective Toxicity of Rhodamine 123 in Carcinoma Cells in Vitro
[CANCER RESEARCH 43, 716-720, February 1983] 0008-5472/83/0043-0000502.00 Selective Toxicity of Rhodamine 123 in Carcinoma Cells in Vitro Theodore J. Lampidis, 1 Samuel D. Bernal, 2 lan C. Summerhayes, and Lan Bo Chen a Sidney Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts 02115 ABSTRACT ing appears to be unique, since chemotherapeutic agents currently in clinical use have not been shown to be particularly The study of mitochondria in situ has recently been facilitated selective between tumorigenic and normal cells in vitro. through the use of rhodamine 123, a mitochondrial-specific fluorescent dye. It has been found to be nontoxic when applied MATERIALS AND METHODS for short periods to a variety of cell types and has thus become an invaluable tool for examining mitochondrial morphology and Cell Cultures. All cell types and cell lines were grown in Dulbecco's function in the intact living cell. In this report, however, we modified Eagle's medium supplemented with 10% calf serum (M. A. demonstrate that with continuous exposure, rhodamine 123 Bioproducts, Walkersville, Md.) at 37 ~ with 5% CO2. The following cell selectively kills carcinoma as compared to normal epithelial lines were obtained from the American Type Culture Collection: cells grown in vitro. At doses of rhodamine 123 which were CCL 105, CCL 15, CRL 1550, CRL 1420, CCL51, CCL77, CCL185, toxic to carcinoma cells, the conversion of mitochondrial-spe- CCL 34, PtK-1, CV-1, and BSC-I. The human bladder carcinoma line cific to cytoplasmic-nonspecific localization of the drug was (E J) was provided by Dr.