Fluorescence Decay Characteristics of the Green Emission From CaF2:Ho3+

Variations in the decay times of the characteristic green emissions at 522.7, 551.3, 549.6, 547.6, 542.2, 540.2, 535.9 and 533.5nm from CaF2 :H03+ with concentration are studied at RT and LNT. A pulsed N2 laser beam of power density 1.5 MW cm-2 is used for the excitation. Temperature dependent concentration quenching of the decay times are observed for all the emission bands. But an increase in the decay time due to the reabsorption process is also observed for a few of the above bands.

Nd:Glass laser-induced damage to thin films, two-photon excited fluorescence and plasma studies

Laser-induced damage is the principal limiting constraint in the design and operation of high-power laser systems used in fusion and other high-energy laser applications. Therefore, an understanding of the mechanisms which cause the radiation damage to the components employed in building a laser and a knowledge of the damage threshold of these materials are of great importance in designing a laser system and to operate it without appreciable degradation in performance. This thesis, even though covers three distinct problems for investigations using a dye Q-switched multimode Nd:glass laser operating at 1062 nm and emitting 25 ns (FWHM) pulses, lays its main thrust on damage threshold studies on thin films. Using the same glass laser two-photon excited fluorescence in rhodamine 6G and generation and characterisation of a carbon plasma have also been carried out. The thesis is presented in seven chapters.

Nitrogen laser excited fluorescence of some rare earth doped alkaline earth fluorides

The present study is mainly concéntrated on the visible fluorescence of Ho3+ ,nd 3+ and Er 3+rare earths in alkaline earth fluoride hosts(caF2,srF2,BaF2) using a nitrogen laser excitation. A nitrogen laser was fabricated and its parametric studies were first carried out.

NIR Spectroscopic and Laser Induced Fluorescence studies of some organic molecules

Vibrational overtone spectroscopy of molecules containing X-H oscillators (X = C, N, O...) has become an effective tool for the study of molecular structure, dynamics, inter and intramolecular interactions, conformational aspects and substituent effects in aliphatic and aromatic compounds. In the present work, the author studied the NIR overtone spectra of some liquid phase organic compounds. The analysis of the CH, NH and OH overtones yielded important structural information about these systems. In an attempt to get information on electronic energy levels, we studied the pulsed Nd:YAG laser induced fluorescence spectra of certain organic compounds. The pulsed laser Raman spectra of some organic compounds are also studied. The novel high resolution technique of near infrared tunable diode laser absorption spectroscopy (TDLAS) is used to record the rotational structure of the second OH overtone spectrum of 2-propanol. The spectral features corresponding to the different molecular conformations could be identified from the high resolution spectrum. The whole work described in this thesis is divided into five chapters.

Nd:Glass Laser-Induced Damage To Thin Films, Two-Photon Excited Fluorescence And Plasma Studies

Laser-induced damage is the principal limiting constraint in the design and operation of high-power laser systems used in fusion and other high-energy laser applications. Therefore, an understanding of the mechanisms which cause the radiation damage to the components employed in building a laser and a knowledge of the damage threshold of these materials are of great importance in designing a laser system and to operate it without appreciable degradation in performance. This thesis, even though covers three distinct problems for investigations using a dye Q-switched multimode Nd:glass laser operating at 1062 nm and emitting 25 ns (FWHM) pulses, lays its main thrust on damage threshold studies on thin films. Using the same glass laser two-photon excited fluorescence in rhodamine 6G and generation and characterisation of a carbon plasma have also been carried out.

“Investigations on Vibrational Overtones in the NIR region and Laser Induced Fluorescence in some selected compounds”

The central theme of this research concerns the study of vibrationally excited molecules. We have used the local mode description of such vibrational states, and this -model has now gained general acceptance. A central feature of the model is the Wloealizafion of vibrational energy. A study of these high—energy localized states provides example, becauseof this localization, overtone spectra, which measure the absorption of T vibrational energy, are extremely sensitive to the properties of X-H bonds. We also use -overtone spectra to study the conformation of molecules, i.e., the relative internal orientation of their bonds. The thesis comprises six chapters

Design and study of self-assembled functional organic and hybrid systems for biological applications

The focus of self-assembly as a strategy for the synthesis has been confined largely to molecules, because of the importance of manipulating the structure of matter at the molecular scale. We have investigated the influence of temperature and pH, in addition to the concentration of the capping agent used for the formation of the nano-bio conjugates. For example, the formation of the narrower size distribution of the nanoparticles was observed with the increase in the concentration of the protein, which supports the fact that γ-globulin acts both as a controller of nucleation as well as stabiliser. As analyzed through various photophysical, biophysical and microscopic techniques such as TEM, AFM, C-AFM, SEM, DLS, OPM, CD and FTIR, we observed that the initial photoactivation of γ-globulin at pH 12 for 3 h resulted in small protein fibres of ca. Further irradiation for 24 h, led to the formation of selfassembled long fibres of the protein of ca. 5-6 nm and observation of surface plasmon resonance band at around 520 nm with the concomitant quenching of luminescence intensity at 680 nm. The observation of light triggered self-assembly of the protein and its effect on controlling the fate of the anchored nanoparticles can be compared with the naturally occurring process such as photomorphogenesis.Furthermore,our approach offers a way to understand the role played by the self-assembly of the protein in ordering and knock out of the metal nanoparticles and also in the design of nano-biohybrid materials for medicinal and optoelectronic applications. Investigation of the potential applications of NIR absorbing and water soluble squaraine dyes 1-3 for protein labeling and anti-amyloid agents forms the subject matter of the third chapter of the thesis. The study of their interactions with various proteins revealed that 1-3 showed unique interactions towards serum albumins as well as lysozyme. 69%, 71% and 49% in the absorption spectra as well as significant quenching in the fluorescence intensity of the dyes 1-3, respectively. Half-reciprocal analysis of the absorption data and isothermal titration calorimetric (ITC) analysis of the titration experiments gave a 1:1 stoichiometry for the complexes formed between the lysozyme and squaraine dyes with association constants (Kass) in the range 104-105 M-1. We have determined the changes in the free energy (ΔG) for the complex formation and the values are found to be -30.78, -32.31 and -28.58 kJmol-1, respectively for the dyes 1, 2 and 3. Furthermore, we have observed a strong induced CD (ICD) signal corresponding to the squaraine chromophore in the case of the halogenated squaraine dyes 2 and 3 at 636 and 637 nm confirming the complex formation in these cases. To understand the nature of interaction of the squaraine dyes 1-3 with lysozyme, we have investigated the interaction of dyes 1-3 with different amino acids. These results indicated that the dyes 1-3 showed significant interactions with cysteine and glutamic acid which are present in the side chains of lysozyme. In addition the temperature dependent studies have revealed that the interaction of the dye and the lysozyme are irreversible. Furthermore, we have investigated the interactions of these NIR dyes 1-3 with β- amyloid fibres derived from lysozyme to evaluate their potential as inhibitors of this biologically important protein aggregation. These β-amyloid fibrils were insoluble protein aggregates that have been associated with a range of neurodegenerative diseases, including Huntington, Alzheimer’s, Parkinson’s, and Creutzfeldt-Jakob diseases. We have synthesized amyloid fibres from lysozyme through its incubation in acidic solution below pH 4 and by allowing to form amyloid fibres at elevated temperature. To quantify the binding affinities of the squaraine dyes 1-3 with β-amyloids, we have carried out the isothermal titration calorimetric (ITC) measurements. The association constants were determined and are found to be 1.2 × 105, 3.6× 105 and 3.2 × 105 M-1 for the dyes, 1-3, respectively. To gain more insights into the amyloid inhibiting nature of the squaraine dyes under investigations, we have carried out thioflavin assay, CD, isothermal titration calorimetry and microscopic analysis. The addition of the dyes 1-3 (5μM) led to the complete quenching in the apparent thioflavin fluorescence, thereby indicating the destabilization of β-amyloid fibres in the presence of the squaraine dyes. Further, the inhibition of the amyloid fibres by the squaraine dyes 1-3, has been evidenced though the DLS, TEM AFM and SAED, wherein we observed the complete destabilization of the amyloid fibre and transformation of the fibre into spherical particles of ca. These results demonstrate the fact that the squaraine dyes 1-3 can act as protein labeling agents as well as the inhibitors of the protein amyloidogenesis. The last chapter of the thesis describes the synthesis and investigation of selfassembly as well as bio-imaging aspects of a few novel tetraphenylethene conjugates 4-6.Expectedly, these conjugates showed significant solvatochromism and exhibited a hypsochromic shift (negative solvatochromism) as the solvent polarity increased, and these observations were justified though theoretical studies employing the B3LYP/6-31g method. We have investigated the self-assembly properties of these D-A conjugates though variation in the percentage of water in acetonitrile solution due to the formation of nanoaggregates. Further the contour map of the observed fluorescence intensity as a function of the fluorescence excitation and emission wavelength confirmed the formation of J-type aggregates in these cases. To have a better understanding of the type of self-assemblies formed from the TPE conjugates 4-6, we have carried out the morphological analysis through various microscopic techniques such as DLS, SEM and TEM. 70%, we observed rod shape architectures having ~ 780 nm in diameter and ~ 12 μM in length as evidenced through TEM and SEM analysis. We have made similar observations with the dodecyl conjugate 5 at ca. 70% and 50% water/acetonitrile mixtures, the aggregates formed from 4 and 5 were found to be highly crystalline and such structures were transformed to amorphous nature as the water fraction was increased to 99%. To evaluate the potential of the conjugate as bio-imaging agents, we have carried out their in vitro cytotoxicity and cellular uptake studies though MTT assay, flow cytometric and confocal laser scanning microscopic techniques. Thus nanoparticle of these conjugates which exhibited efficient emission, large stoke shift, good stability, biocompatibility and excellent cellular imaging properties can have potential applications for tracking cells as well as in cell-based therapies. In summary we have synthesized novel functional organic chromophores and have studied systematic investigation of self-assembly of these synthetic and biological building blocks under a variety of conditions. The investigation of interaction of water soluble NIR squaraine dyes with lysozyme indicates that these dyes can act as the protein labeling agents and the efficiency of inhibition of β-amyloid indicate, thereby their potential as anti-amyloid agents.

Novel Electrochemical and Fluorescence Sensors for Food Additives and Neurotransmitters

There is an enormous demand for chemical sensors in many areas and disciplines including chemistry, biology, clinical analysis, environmental science. Chemical sensing refers to the continuous monitoring of the presence of chemical species and is a rapidly developing field of science and technology. They are analytical devices which transform chemical information generating from a reaction of the analyte into an measurable signal. Due to their high selectivity, sensitivity, fast response and low cost, electrochemical and fluorescent sensors have attracted great interest among the researchers in various fields. Development of four electrochemical sensors and three fluorescent sensors for food additives and neurotransmitters are presented in the thesis. Based on the excellent properties of multi walled carbon nanotube (MWCNT), poly (L-cysteine) and gold nanoparticles (AuNP) four voltammetric sensors were developed for various food additives like propyl gallate, allura red and sunset yellow. Nanosized fluorescent probes including gold nanoclusters (AuNCs) and CdS quantum dots (QDs) were used for the fluorescent sensing of butylated hydroxyanisole, dopamine and norepinephrine. A total of seven sensors including four electrochemical sensors and three fluorescence sensors have been developed for food additives and neurotransmitters.

Fluorescence studies on the 3d-ionization threshold range in krypton

Krypton atoms were excited by photons in the energy range from the threshold for photoionization of the 3d-electrons up to 120 eV. and the fluorescence radiation in the spectral range from 780 to 965 A was observed and analyzed. Cross sections for the population of excited states in KrIII with at least one 4s-hole resulting from an Auger transition as the first decay step and for KrII satellites were determined. The energy dependence of the 3d-ionization cross section in the 3d{_5/2}- and the 3d{_3/2}-threshold range was derived from the experimental data. The cross sections for production of KrII states were found to follow the energy dependence of the 3d-cross sections.

Absolute Kr 4s-electron photoionization cross sections between 30 and 90 eV measured by photon-induced fluorescence spectroscopy (PIFS)

Absolute Kr 4s-electron photoionization cross sections as a function of the exciting-photon energy between 30 and 90 eV were measured by photon-induced fluorescence spectroscopy (PIFS). The measurements were compared with available experimental data and theoretical calculations.

Detecting ultrafast interatomic electronic processes in media by fluorescence

Interatomic coulombic decay (ICD), a radiationless transition in weakly bonded systems, such as solutes or van der Waals bound aggregates, is an effective source for electrons of low kinetic energy. So far, the ICD processes could only be probed in ultra-high vacuum by using electron and/or ion spectroscopy. Here we show that resonant ICD processes can also be detected by measuring the subsequently emitted characteristic fluorescence radiation, which makes their study in dense media possible.

Surface Reflectance Recognition and Real-World Illumination Statistics

Humans distinguish materials such as metal, plastic, and paper effortlessly at a glance. Traditional computer vision systems cannot solve this problem at all. Recognizing surface reflectance properties from a single photograph is difficult because the observed image depends heavily on the amount of light incident from every direction. A mirrored sphere, for example, produces a different image in every environment. To make matters worse, two surfaces with different reflectance properties could produce identical images. The mirrored sphere simply reflects its surroundings, so in the right artificial setting, it could mimic the appearance of a matte ping-pong ball. Yet, humans possess an intuitive sense of what materials typically "look like" in the real world. This thesis develops computational algorithms with a similar ability to recognize reflectance properties from photographs under unknown, real-world illumination conditions. Real-world illumination is complex, with light typically incident on a surface from every direction. We find, however, that real-world illumination patterns are not arbitrary. They exhibit highly predictable spatial structure, which we describe largely in the wavelet domain. Although they differ in several respects from the typical photographs, illumination patterns share much of the regularity described in the natural image statistics literature. These properties of real-world illumination lead to predictable image statistics for a surface with given reflectance properties. We construct a system that classifies a surface according to its reflectance from a single photograph under unknown illuminination. Our algorithm learns relationships between surface reflectance and certain statistics computed from the observed image. Like the human visual system, we solve the otherwise underconstrained inverse problem of reflectance estimation by taking advantage of the statistical regularity of illumination. For surfaces with homogeneous reflectance properties and known geometry, our system rivals human performance.

Fluorescence Assay for Polymerase Arrival Rates

To engineer complex synthetic biological systems will require modular design, assembly, and characterization strategies. The RNA polymerase arrival rate (PAR) is defined to be the rate that RNA polymerases arrive at a specified location on the DNA. Designing and characterizing biological modules in terms of RNA polymerase arrival rates provides for many advantages in the construction and modeling of biological systems. PARMESAN is an in vitro method for measuring polymerase arrival rates using pyrrolo-dC, a fluorescent DNA base that can substitute for cytosine. Pyrrolo-dC shows a detectable fluorescence difference when in single-stranded versus double-stranded DNA. During transcription, RNA polymerase separates the two strands of DNA, leading to a change in the fluorescence of pyrrolo-dC. By incorporating pyrrolo-dC at specific locations in the DNA, fluorescence changes can be taken as a direct measurement of the polymerase arrival rate.

Perceiving Illumination Inconsistencies in Scenes

The human visual system is adept at detecting and encoding statistical regularities in its spatio-temporal environment. Here we report an unexpected failure of this ability in the context of perceiving inconsistencies in illumination distributions across a scene. Contrary to predictions from previous studies [Enns and Rensink, 1990; Sun and Perona, 1996a, 1996b, 1997], we find that the visual system displays a remarkable lack of sensitivity to illumination inconsistencies, both in experimental stimuli and in images of real scenes. Our results allow us to draw inferences regarding how the visual system encodes illumination distributions across scenes. Specifically, they suggest that the visual system does not verify the global consistency of locally derived estimates of illumination direction.

Synthesis, Optical properties and Applications of Water Soluble Conjugated PPPs for Biosensors

In recent years, application of fluorescent conjugated polymers to sense chemical and biological analytes has received much attention owing to its technological significance. Water soluble conjugated polymers are interesting towards the developing sensors for biomolecules. In this present contribution, we describe the syntheses and characterization of a series of water soluble conjugated polymers with sulfonic acid groups in the side chain. Such anionic conjugated polymers are designed to interact with biomolecules such as cytochrome-C. All polymers are water soluble and showed strong blue emission. Significant quenching of the fluorescence from our functionalized PPP was observed upon addition of viologen derivatives or cytochrome -C.