Relevant bibliographies by topics / Two-photon excitation fluorescence of proteins

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  1. Journal articles
  2. Dissertations / Theses
  3. Book chapters
  4. Conference papers
  5. Reports

Academic literature on the topic 'Two-photon excitation fluorescence of proteins'

Author: Grafiati
Published: 4 June 2021
Last updated: 8 February 2022

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Journal articles on the topic "Two-photon excitation fluorescence of proteins"

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Diaspro, Alberto, Giuseppe Chirico, and Maddalena Collini. "Two-photon fluorescence excitation and related techniques in biological microscopy." Quarterly Reviews of Biophysics 38, no. 2 (May 2005): 97–166. http://dx.doi.org/10.1017/s0033583505004129.

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1. Introduction 982. Historical background of two-photon effects 992.1 2PE 1002.2 Harmonic generation 1002.3 Fluorescence correlation spectroscopy 1003. Basic principles of two-photon excitation of fluorescent molecules and implications for microscopy and spectroscopy 1013.1 General considerations 1013.2 Fluorescence intensity under the 2PE condition 1033.3 Optical consequences of 2PE 1043.4 Saturation effects in 2PE 1083.5 Fluorescence correlation spectroscopy 1093.5.1 Autocorrelation analysis 1103.5.2 Photon-counting histogram analysis 1124. Two-photon-excited probes 1155. Design considerations for a 2PE fluorescence microscope 1195.1 General aspects 1195.2 Descanned and non-descanned 2PE imaging 1215.3 Lens objectives and pulse broadening 1225.4 Laser sources 1255.5 Example of a practical realization 1276. Applications 1346.1 Biological applications of 2PE 1346.1.1 Brain images 1346.1.2 Applications on the kidney 1396.1.3 Mammalian embryos 1396.1.4 Applications to immuno-response 1416.1.5 Myocytes 1416.1.6 Retina 1426.1.7 DNA imaging 1436.1.8 FISH applications 1446.2 2PE imaging of single molecules 1446.3 FCS applications 1486.4 Signals from nonlinear interactions 1517. Conclusions 1538. Acknowledgements 1549. References 155This review is concerned with two-photon excited fluorescence microscopy (2PE) and related techniques, which are probably the most important advance in optical microscopy of biological specimens since the introduction of confocal imaging. The advent of 2PE on the scene allowed the design and performance of many unimaginable biological studies from the single cell to the tissue level, and even to whole animals, at a resolution ranging from the classical hundreds of nanometres to the single molecule size. Moreover, 2PE enabled long-term imaging of in vivo biological specimens, image generation from deeper tissue depth, and higher signal-to-noise images compared to wide-field and confocal schemes. However, due to the fact that up to this time 2PE can only be considered to be in its infancy, the advantages over other techniques are still being evaluated. Here, after a brief historical introduction, we focus on the basic principles of 2PE including fluorescence correlation spectroscopy. The major advantages and drawbacks of 2PE-based experimental approaches are discussed and compared to the conventional single-photon excitation cases. In particular we deal with the fluorescence brightness of most used dyes and proteins under 2PE conditions, on the optical consequences of 2PE, and the saturation effects in 2PE that mostly limit the fluorescence output. A complete section is devoted to the discussion of 2PE of fluorescent probes. We then offer a description of the central experimental issues, namely: choice of microscope objectives, two-photon excitable dyes and fluorescent proteins, choice of laser sources, and effect of the optics on 2PE sensitivity. An inevitably partial, but vast, overview of the applications and a large and up-to-date bibliography terminate the review. As a conclusive comment, we believe that 2PE and related techniques can be considered as a mainstay of the modern biophysical research milieu and a bright perspective in optical microscopy.
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Paul, Uchenna P., Li, Milton L. Lee, and Paul B. Farnsworth. "Compact Detector for Proteins Based on Two-Photon Excitation of Native Fluorescence." Analytical Chemistry 77, no. 11 (June 2005): 3690–93. http://dx.doi.org/10.1021/ac048161z.

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Wan, H., C. Soeller, D. R. Garrod, C. Robinson, and M. B. Cannell. "Quantitative Immunocytochemistry of Proteins Using 2- Photon Microscopy and Digital Image Analysis." Microscopy and Microanalysis 4, S2 (July 1998): 416–17. http://dx.doi.org/10.1017/s1431927600022200.

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The two photon microscope provides optical sectioning of fluorescent specimens with a resolution comparable to that obtained in confocal microscopy (see refs 2,3). However, the excited volume in 2-photon microscopy is limited to the focal volume (unlike conventional fluorescence microscopy where excitation occurs throughout the specimen). This means that photodamage is limited to the plane of section being examined. Thus, the light emitted from each point in the specimen depends on the amount of fluorochrome present without the problem of prior illumination (of other planes within the specimen) reducing the photon yield so a better signal to noise ratio can be obtained when examination of multiple image planes is needed. Since 2-photon excitation spectra are wide and chromatic aberration is eliminated (because the emitted light does not have to be focused on a pinhole), it is possible to excite several fluorochromes simultaneously and map their positions with high accuracy.
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Botchway, Stanley W., Ignasi Barba, Randolf Jordan, Rebecca Harmston, Peter M. Haggie, Simon-Peter Williams, Alexandra M. Fulton, Anthony W. Parker, and Kevin M. Brindle. "A novel method for observing proteins in vivo using a small fluorescent label and multiphoton imaging." Biochemical Journal 390, no. 3 (September 5, 2005): 787–90. http://dx.doi.org/10.1042/bj20050648.

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A novel method for the fluorescence detection of proteins in cells is described in the present study. Proteins are labelled by the selective biosynthetic incorporation of 5-hydroxytryptophan and the label is detected via selective two-photon excitation of the hydroxyindole and detection of its fluorescence emission at 340 nm. The method is demonstrated in this paper with images of a labelled protein in yeast cells.
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Kierdaszuk, Borys, Ignacy Gryczynski, Anna Modrak-Wojcik, Agnieszka Bzowska, David Shugar, and Joseph R. Lakowicz. "FLUORESCENCE OF TYROSINE AND TRYPTOPHAN IN PROTEINS USING ONE- AND TWO-PHOTON EXCITATION." Photochemistry and Photobiology 61, no. 4 (April 1995): 319–24. http://dx.doi.org/10.1111/j.1751-1097.1995.tb08615.x.

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Chirico, G., A. Diaspro, F. Cannone, M. Collini, S. Bologna, V. Pellegrini, and F. Beltram. "Selective Fluorescence Recovery after Bleaching of Single E2GFP Proteins Induced by Two-Photon Excitation." ChemPhysChem 6, no. 2 (February 11, 2005): 328–35. http://dx.doi.org/10.1002/cphc.200400318.

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Wang, Ke, Tzu-Ming Liu, Juwell Wu, Nicholas G. Horton, Charles P. Lin, and Chris Xu. "Three-color femtosecond source for simultaneous excitation of three fluorescent proteins in two-photon fluorescence microscopy." Biomedical Optics Express 3, no. 9 (July 31, 2012): 1972. http://dx.doi.org/10.1364/boe.3.001972.

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Cannell, M. B., and C. Soeller. "Optical Sectioning in Fluorescence Microscopy by Confocal and 2-Photon Molecular Excitation Techniques." Microscopy Today 5, no. 8 (October 1997): 12–15. http://dx.doi.org/10.1017/s1551929500056741.

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Fluorescence microscopy has proved to be an invaluable tool for biomedical science since it is possible to visualise small quantities of labeled materials (such as intracellular ions and proteins) in both fixed and living cells, However, the conventional wide field fluorescence microscope suffers from the disadvantage that objects outside the focal plane also fluoresce (in response to the excitation light) and this leads to a marked loss of contrast for objects in the focal plane, This is especially a problem when the fluorescent probe is distributed throughout the thickness of the cell and the cell is thicker than about 1 µm. The confocal microscope overcomes this problem by illuminating the preparation with a point source of excitation light and limiting the collection of light with a pinhole that is confocal with the illumination source.
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Cannell, M. B., and C. Soeller. "High Resolution Imaging Using Confocal and 2-photon Molecular Excitation Microscopy." Microscopy Today 8, no. 5 (June 2000): 20–26. http://dx.doi.org/10.1017/s1551929500065196.

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Fluorescence microscopy has proved to be an invaluable tool for biomedical science since it is possible to visualise small quantities of labelled materials (such as intracellular ions and proteins) in both fixed and living cells. However, the conventional wide field fluorescence microscope suffers from the disadvantage that objects outside the focal plane also fluoresce (in response to the excitation light) and this leads to a marked loss of contrast for objects in the focal plane. This is especially a problem when the fluorescent probe is distributed throughout the thickness of the cell and the cell is thicker than about 1 μm. The confocal microscope overcomes this problem by illuminating the preparation with a point source of excitation light and limiting the collection of light with a pinhole that is confocal with the illumination source. This converts the microscope from an imaging system to a point detector and images are produced by scanning the illuminating and detecting point over the specimen to build an image (in much the same way that a television set produces an image). The basic idea behind the confocal system is shown in Figure 1, and it should be noted that light from points outside the focal plane is defocused at the pinhole and so does not pass through the pinhole efficiently.
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Tao, Wen, Michael Rubart, Jennifer Ryan, Xiao Xiao, Chunping Qiao, Takashi Hato, Michael W. Davidson, Kenneth W. Dunn, and Richard N. Day. "A practical method for monitoring FRET-based biosensors in living animals using two-photon microscopy." American Journal of Physiology-Cell Physiology 309, no. 11 (December 1, 2015): C724—C735. http://dx.doi.org/10.1152/ajpcell.00182.2015.

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The commercial availability of multiphoton microscope systems has nurtured the growth of intravital microscopy as a powerful technique for evaluating cell biology in the relevant context of living animals. In parallel, new fluorescent protein (FP) biosensors have become available that enable studies of the function of a wide range of proteins in living cells. Biosensor probes that exploit Förster resonance energy transfer (FRET) are among the most sensitive indicators of an array of cellular processes. However, differences between one-photon and two-photon excitation (2PE) microscopy are such that measuring FRET by 2PE in the intravital setting remains challenging. Here, we describe an approach that simplifies the use of FRET-based biosensors in intravital 2PE microscopy. Based on a systematic comparison of many different FPs, we identified the monomeric (m) FPs mTurquoise and mVenus as particularly well suited for intravital 2PE FRET studies, enabling the ratiometric measurements from linked FRET probes using a pair of experimental images collected simultaneously. The behavior of the FPs is validated by fluorescence lifetime and sensitized emission measurements of a set of FRET standards. The approach is demonstrated using a modified version of the AKAR protein kinase A biosensor, first in cells in culture, and then in hepatocytes in the liver of living mice. The approach is compatible with the most common 2PE microscope configurations and should be applicable to a variety of different FRET probes.
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Dissertations / Theses on the topic "Two-photon excitation fluorescence of proteins"

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Li, Li. "Detection of Proteins by Two-Photon Excitation of Native Fluorescence." Diss., CLICK HERE for online access, 2006. http://contentdm.lib.byu.edu/ETD/image/etd1548.pdf.

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Paul, Uchenna Prince. "Fluorescence Detectors for Proteins and Toxic Heavy Metals." Diss., CLICK HERE for online access, 2004. http://contentdm.lib.byu.edu/ETD/image/etd416.pdf.

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Mikaelsson, Therese. "Electronic Energy Migration/Transfer as a Tool to Explore Biomacromolecular Structures." Doctoral thesis, Umeå universitet, Kemiska institutionen, 2014. http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-86794.

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Fluorescence-based techniques are widely used in bioscience, offering a high sensitivity and versatility. In this work, fluorescence electronic energy migration/ transfer is applied to measure intramolecular distances in two types of systems and under various conditions. The main part of the thesis utilizes the process of donor-acceptor energy transfer to probe distances within the ribosomal protein S16. Proteins are essential to all organisms. Therefore, it is of great interest to study protein structure and function in order to understand and prevent protein malfunction. Moreover, it is also important to try to study the proteins in an environment which resembles its natural habitat. Here two protein homologs were investigated; S16Thermo and S16Meso, isolated from a hyperthemophilic bacterium and a mesophilic bacterium, respectively. It was concluded that the chemically induced unfolded state ensemble of S16Thermo is more compact than the corresponding ensemble of S16Meso. This unfolded state compaction may be one reason for the increased thermal stability of S16Thermo as compared to S16Meso. The unfolded state of S16 was also studied under highly crowded conditions, mimicking the environment found in cells. It appears that a high degree of crowding, induced by 200 mg/mL dextran 20, forces the unfolded state ensemble of S16Thermo to become even more compact. Further, intramolecular distances in the folded state of five S16 mutants were investigated upon increasing amounts of dextran 20. We found that the probed distances in S16Thermo are unaffected by increasing degree of crowding. However, S16Meso shows decreasing intramolecular distances for all three studied variants, up to 100 mg/mL dextran. At higher concentrations, the change in distance becomes anisotropic. This suggests that marginally stable proteins like s16Meso may respond to macromolecular crowding by fine-tuning its structure. More stable proteins like S16Thermo however, show no structural change upon increasing degree of crowding. We also investigated the possibility of local specific interactions between the protein and crowding agent, by means of fluorescence quenching experiments. Upon increasing amounts of a tyrosine labelled dextran, a diverse pattern of fluorescence quantum yield and lifetime suggests that specific, local protein-crowder interactions may occur. In a second studied system, electronic energy migration between two donor-groups, separated by a rigid steroid, was studied by two-photon excitation depolarization experiments. Data were analysed by using recent advances, based on the extended Förster theory, which yield a reasonable value of the distance between the two interacting donor-groups. To the best of our knowledge, this is the first quantitative analysis of energy migration data, obtained from two-photon excited fluorescence.
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DeArmond, Fredrick Michael. "Two-Photon Excitation, Fluorescence Microscopy, and Quantitative Measurement of Two-Photon Absorption Cross Sections." PDXScholar, 2017. https://pdxscholar.library.pdx.edu/open_access_etds/4036.

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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 an increased demand for robust non-linear fluorescent markers, and characterization of the associated tool set. These factors have led to an experimental setup to allow a systematized approach for identifying and characterizing properties of fluorescent probes in the hopes that the tool set will provide researchers with additional information to guide their efforts in developing novel fluorophores suitable for use in advanced optical microscopy techniques as well as identifying trends for their synthesis. Hardware was setup around a software control system previously developed [1]. Three experimental tool sets were set up, characterized, and applied over the course of this work. These tools include scanning multiphoton fluorescence microscope with single molecule sensitivity, an interferometric autocorrelator for precise determination of the bandwidth and pulse width of the ultrafast Titanium Sapphire excitation source, and a simplified fluorescence microscope for the measurement of two-photon absorption cross sections. Resulting values for two-photon absorption cross sections and two-photon absorption action cross sections for two standardized fluorophores, four commercially available fluorophores, and ten novel fluorophores are presented as well as absorption and emission spectra.
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Ryderfors, Linus. "Two-Photon Excited Fluorescence Depolarisation : Experimental and Theoretical Development." Doctoral thesis, Uppsala University, Department of Photochemistry and Molecular Science, 2008. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-9285.

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We have studied fundamental aspects of time-resolved two-photon excited fluorescence depolarisation. The thesis presents experimental as well as theoretical progress. We show that a multi-photon induced instrumental response function obtained from a suspension of gold nanoparticles is appropriate for the analysis of two-photon excited fluorescence decays obtained using time-correlated single photon counting detection. Theoretical expressions have been derived for the fluorescence anisotropy decay obtained upon two-photon excitation of various molecular systems in liquid solutions: a) an anisotropic rigid rotor that undergoes rotational diffusion in the presence of ultrafast unresolved restricted reorientations, e.g. librations. b) a molecular group covalently attached to a stationary macromolecule, and undergoing local reorientation in a uniaxial ordering potential. A new approach to the analysis of two-photon excited fluorescence depolarisation experiments was developed, which combines data obtained by using linearly and circularly polarised excitation light, in a global manner. In the analysis, knowledge about unresolved reorientations was obtained from one-photon excitation studies of the corresponding systems. By means of this procedure it has been possible to obtain quantitative information about the molecular two-photon absorption tensor for perylene and two of its derivatives. Thereby the symmetry of the final excited and intermediate vibronic states could be assigned. The analysis reveals that the two-photon transition studied with the 800 nm laser exhibits mixed character. An important finding from the experiments was that the two-photon absorption tensor appears to be solvent dependent. Furthermore, the thesis presents the first theoretical treatment of two-photon excited donor-donor energy migration in the presence of molecular reorientation and which applies the extended Förster theory. Explicit expressions for molecules that belong to the point groups D2h, D2 and C2v are given. Preliminary experiments are finally also reported on a two-photon excited donor-donor energy migration system consisting of a bisanthryl-bisteroid.

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Ogden, Melinda Anne. "Two-photon total internal reflection microscopy for imaging live cells with high background fluorescence." Thesis, Georgia Institute of Technology, 2009. http://hdl.handle.net/1853/34786.

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Fluorescence microscopy allows for spatial and temporal resolution of systems which are inherently fluorescent or which can be selectively labeled with fluorescent molecules. Temporal resolution is crucial for imaging real time processes in living samples. A common problem in fluorescence microscopy of biological samples is autofluorescence, fluorescence inherent to the system, which interferes with detection of fluorescence of interest by decreasing the signal to noise ratio. Two current methods for improved imaging against autofluorescence are two-photon excitation and total internal reflection microscopy. Two-photon excitation occurs when two longer wavelength photons are absorbed quasi-simultaneously by a single fluorophore. For this to take place there must be a photon density on the order of 1030 photons/(cm2)(s), which is achieved through use of a femtosecond pulsed laser and a high magnification microscope objective. Two-photon excitation then only occurs at the focal spot, significantly reducing the focal volume and therefore background autofluorescence. The second method, total internal reflection, is based on evanescent wave excitation, which decreases exponentially in intensity away from the imaging surface. This allows for excitation of a thin (~200 nm) slice of a sample. Since only a narrow region of interest is excited, an optical slice can be imaged, decreasing excitation of out-of-focus autofluorescence, and increasing the signal to noise ratio. By coupling total internal reflection with two-photon excitation, an entire cell can be imaged while still maintaining the use of lower energy photons to irradiate the sample and achieve two-photon excitation along the length traveled by the evanescent wave. This system allows for more sensitive detection of fluorescence of interest from biological systems as a result of a significant decrease in excitation volume and therefore a decrease in autofluorescence signal. In the two-photon total internal reflection microscopy setup detailed in this work, an excitation area of 20 μm by 30 μm is achieved, and used to image FITC-stained actin filaments in BS-C-1 cells
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Takasaki, Kevin Takao. "Development and Application of Two-Photon Excitation Stimulated Emission Depletion Microscopy for Superresolution Fluorescence Imaging in Thick Tissue." Thesis, Harvard University, 2013. http://dissertations.umi.com/gsas.harvard:10976.

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Two-photon laser scanning microscopy (2PLSM) allows fluorescence imaging in thick biological samples where absorption and scattering typically degrade resolution and signal collection of 1-photon imaging approaches. The spatial resolution of conventional 2PLSM is limited by diffraction, and the near-infrared wavelengths used for excitation in 2PLSM preclude the accurate imaging of many small subcellular features of neurons. Stimulated emission depletion (STED) microscopy is a superresolution imaging modality which overcomes the resolution limit imposed by diffraction and allows fluorescence imaging of nanoscale features. In this thesis, I describe the development of 2PLSM combined with STED microscopy for superresolution fluorescence imaging of neurons embedded in thick tissue. Furthermore, I describe the application of this method to studying the biophysics connecting synaptic structure and function in dendritic spines.
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Fisher, Rachel Sarah. "Photophysical characterisation of novel fluorescent base analogues." Thesis, University of Edinburgh, 2018. http://hdl.handle.net/1842/31271.

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Fluorescent nucleic acid base analogues (FBAs) are an important class of molecule used to study the structure and dynamics of DNA and RNA. These base analogues are molecules with structures that resemble one of the natural bases but which, unlike the natural bases, have high fluorescence quantum yields. 2-Aminopurine (2AP) has long been the most widely used fluorescent base analogue and is one of the few base analogues commercially available. One problem with 2AP is that it undergoes significant quenching when incorporated into DNA: the quantum yield decreases 100 fold from that of the free base, thus becoming too low for use in, for example, single molecule studies. A secondary problem is that the 305 nm absorption peak requires excitation in the UV. A variety of new fluorescent base analogues are being produced, with a view to remedying the deficiencies of 2AP and expanding the current range of use. The first part of this thesis explores the one-photon photophysical properties of several of these novel FBAs. The first of these novel FBAs is the 6-aza-uridine family. These compounds, analogues of uridine, have large Stokes shifts and their absorption and emission spectra are red-shifted in comparison to 2AP; their quantum yields as free bases have been shown to exceed that of 2AP and their environmental sensitivity has been demonstrated. Time-resolved measurements reported in this thesis indicate the presence of multiple emitting species. A density functional theory (DFT) study has been carried out to rationalise these emitting species as rotational isomers. Similar fluorescence lifetime measurements were made on a second class of FBAs, the quadracyclic adenine analogues, qANs; these results also indicated the presence of multiple emitting species. Experimental results show that these FBAs undergo excited-state proton transfer. The final FBA studied in this thesis is pentacyclic adenine, pA. This FBA showed some of the most promising characteristics of all the analogues investigated, such as a high quantum yield in both polar and non-polar solvents. A time-resolved investigation into pA-containing oligonucleotides indicated that in an oligonucleotide pA adopts multiple stacked conformations and its behaviour is highly sequence dependent. Several of these aforementioned fluorescent base analogues have absorption spectra in a region that makes them accessible to two-photon (2P) excitation with a Ti:Sapphire laser. In biological systems, multiphoton excitation has several advantages over one-photon excitation. By avoiding the use of ultraviolet light there is reduced phototoxicity. Out of focus photobleaching and autofluorescence are also minimised which leads to a higher signal-to-background ratio and allows deeper tissue penetration to be achieved. Fluorescent base analogues tend to have small Stokes shifts; this is another problem that can be overcome by using two-photon excitation. To be of potential use in multiphoton microscopy, a FBA must have a high two-photon absorption cross-section and a high two-photon brightness. Previously, the highest two- photon brightness measured for a fluorescent base analogue was less than 2 GM. Amongst the base analogues investigated here, are several that have higher two-photon brightness than ever reported for FBAs; these include pA which is shown to have the highest 2P brightness of a FBA in an oligonucleotide, 1.3 GM, and a member of the 6-azauridine family which as a free base has a 2P brightness of 18 GM. Detection of individual molecules represents the ultimate level of sensitivity and enables details about a molecular system that would otherwise be concealed using conventional ensemble techniques to be revealed. With the improved 2P brightness of the molecules measured in this thesis, it has become feasible to detect single FBA molecules using 2P excitation. To maximise the chance of detection, ultrafast, shaped laser pulses have been used as the excitation source. For the first time, the signal has been high enough and the molecule of interest sufficiently photostable such that 2P fluorescence correlation spectroscopy of a fluorescent base analogue in an oligonucleotide could be measured. In summary, this thesis reports the fluorescence lifetimes and two-photon cross-sections of a series of novel fluorescent base analogues, as well as fluorescence correlation spectroscopy measurements of the most promising candidates.
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Opanasyuk, Oleg. "A new approach to the analyses of fluorescence depolarisation experiments in the presence of electronic energy transport." Doctoral thesis, Umeå universitet, Kemiska institutionen, 2011. http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-50050.

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A new and general procedure is described for a detailed analysis of time-resolved fluorescence depolarisation data in the presence of electronic energy migration. An isotropic ensemble of bifluorophoric molecules (D1-R-D2) has been studied to demonstrate its utility. Intramolecular donor-donor energy migration occurs between the two donor groups (D), which are covalently connected to a rigid linker group (R). These groups undergo restricted reorientational motions with respect to the R group. The analysis of depolarisation data basically involves the search for best-fit parameters which describe the local reorienting motions, the interfluorophore D1-D2 distance, as well as the mutual orientations of the donors. For this, the analysis is partly performed in the Fourier domain and the best-fit parameters are determined by using an approach based on a Genetic Algorithm. The energy migration process has been described by using Monte Carlo simulations and an extended Förster theory. It is found that this theory provides the least time-consuming computational method. Since one-photon and two-photon excited fluorescence experiments can be applied for energy migration studies, a general and unified theoretical formulation is given. To exemplify the developed quantitative approach the depolarisation of the fluorescence in the presence of electronic energy migration within a bis-(9-anthrylmethylphosphonate) bisteroid molecule has been studied by time-resolved two-photon excited fluorescence depolarisation experiments. To solely obtain information about local reorientations of the 9-anthrylmethyl group, also the mono-(9-anthrylmethylphosphonate) bisteroid was studied, which enabled modelling of the ordering potential of the donor. Values of the two-photon absorption tensor components were obtained. To describe the discrepancy between the measured values of the initial anisotropy and fundamental anisotropy predicted by theory the distribution of absorption tensor caused by fast processes have been introduced. An angular parameter of absorption tensor was determined. Reasonable values of the distance between the 9-anthrylmethyl groups, as well as for their mutual orientation were obtained.
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Massin, Julien. "Ingénierie moléculaire pour l'imagerie par microscopie non-linéaire : synthèse et propriétés de nouvelles sondes." Thesis, Lyon, École normale supérieure, 2011. http://www.theses.fr/2011ENSL0701.

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L’objectif de cette thèse est l’élaboration de sondes organiques pour la microscopie optique non-linéaire par fluorescence excitée à deux photons (F2P) et génération de seconde harmonique (GSH). Dans une première partie, cette thèse décrit la synthèse de sondes pour l’imagerie de potentiels de membrane par GSH, comportant un ou plusieurs motifs sucres ainsi que leurs caractérisations spectroscopiques. Les premiers essais en imagerie biologique ont permis de démontrer une bonne affinité des sondes sucres pour la membrane cellulaire et un signal de GSH sur cellule neuronale a pu être observé sur une période de temps allant jusqu'à près de trois heures. La seconde approche a consisté à synthétiser et étudier des chromophores possédant des propriétés de fluorescence à l’état solide pour des applications dans la synthèse de nanoparticules fluorescentes pour l’imagerie biologique. 18 des 21 composés synthétisés ont pu être cristallisés et leur structure résolue par diffraction des rayons X et les propriétés spectroscopique en solution et à l’état solide ont été réalisées. Cette étude a permis de montrer que l’arrangement des molécules les unes par rapport aux autres avait une grande influence sur la fluorescence à l’état solide et donc que les substituants avaient une grande importance. Enfin, cette partie se termine sur les premiers essais effectués pour synthétiser des nanoparticules fluorescentes
The objective of this thesis is the design of new organic probes for nonlinear optical microscopy by two-photon excited fluorescence (TPEF) and second harmonic generation (SHG). In the first part, we describe the synthesis of probes for voltage sensitive imaging by SHG, bearing one or more sugar units and their spectroscopic characterization. The first biological imaging tests have shown good affinity of the probes to the cell membrane and the SHG signal of neuronal cell was observed over a period of nearly three hours. The second part comprises the synthesis and the study of chromophores with solid state fluorescence properties for use in fluorescent nanoparticles for biological imaging. 18 of the 21 compounds synthesized have been crystallized, their crystal structures determined by X-ray diffraction and their spectroscopic properties studied in solution and in the solid state. These studies showed that the arrangement of molecules relative to each had a great influence on the solid state fluorescence and therefore that the substitution was very important. The chapter ends with the first tests of fluorescent nanoparticles synthesis
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Book chapters on the topic "Two-photon excitation fluorescence of proteins"

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Nifosì, R., and V. Tozzini. "One-Photon and Two-Photon Excitation of Fluorescent Proteins." In Springer Series on Fluorescence, 3–40. Berlin, Heidelberg: Springer Berlin Heidelberg, 2011. http://dx.doi.org/10.1007/4243_2011_26.

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Diaspro, Alberto, Paolo Bianchini, Francesca Cella Zanacchi, and Cesare Usai. "Two-Photon Excitation Fluorescence Microscopy." In Encyclopedia of Biophysics, 2667–71. Berlin, Heidelberg: Springer Berlin Heidelberg, 2013. http://dx.doi.org/10.1007/978-3-642-16712-6_829.

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Diaspro, Alberto, Marc Schneider, Paolo Bianchini, Valentina Caorsi, Davide Mazza, Mattia Pesce, Ilaria Testa, Giuseppe Vicidomini, Cesare Usai, and A. Van den Bos. "Two-Photon Excitation Fluorescence Microscopy." In Science of Microscopy, 751–89. New York, NY: Springer New York, 2007. http://dx.doi.org/10.1007/978-0-387-49762-4_11.

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Fahrni, Christoph J. "Fluorescent Probes for Two-Photon Excitation Microscopy." In Reviews in Fluorescence, 249–69. New York, NY: Springer New York, 2009. http://dx.doi.org/10.1007/978-0-387-88722-7_11.

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Johnson, Carey K., and Chaozhi Wan. "Anisotropy Decays Induced by Two-Photon Excitation." In Topics in Fluorescence Spectroscopy, 43–85. Boston, MA: Springer US, 2002. http://dx.doi.org/10.1007/0-306-47070-5_2.

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Kierdaszuk, Borys, Ignacy Gryczynski, and Joseph R. Lakowicz. "Two-Photon Induced Fluorescence of Proteins." In Topics in Fluorescence Spectroscopy, 187–209. Boston, MA: Springer US, 2002. http://dx.doi.org/10.1007/0-306-47070-5_5.

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Piston, David W. "Two-Photon Excitation Microscopy for Three-Dimensional Imaging of Living Intact Tissues." In Fluorescence Microscopy, 203–42. Weinheim, Germany: Wiley-VCH Verlag GmbH & Co. KGaA, 2017. http://dx.doi.org/10.1002/9783527687732.ch6.

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Zhang, Taishi, Tingting Zhao, Peiyan Yuan, and Qing-Hua Xu. "Plasmon-Enhanced Two-Photon Excitation Fluorescence and Biomedical Applications." In Surface Plasmon Enhanced, Coupled and Controlled Fluorescence, 211–25. Hoboken, NJ, USA: John Wiley & Sons, Inc., 2017. http://dx.doi.org/10.1002/9781119325161.ch13.

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Van der Wieb Meer, B., and S. Y. Simon Chen. "Two-Photon Excitation and Anisotropy Decays in Membranes and Oriented Systems." In Topics in Fluorescence Spectroscopy, 145–85. Boston, MA: Springer US, 2002. http://dx.doi.org/10.1007/0-306-47070-5_4.

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Kawano, Hiroyuki, Yasuo Nabekawa, Akira Suda, Yu Oishi, Hideaki Mizuno, Atsushi Miyawaki, and Katsumi Midorikawa. "Adaptive Control of Two-Photon Excitation of Green Fluorescent Protein with Shaped Femtosecond Pulses." In Springer Series in OPTICAL SCIENCES, 449–54. New York, NY: Springer New York, 2004. http://dx.doi.org/10.1007/978-0-387-34756-1_56.

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Conference papers on the topic "Two-photon excitation fluorescence of proteins"

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Wang, Ke, Tzu-Ming Liu, Juwell Wu, Nicholas G. Horton, Charles P. Lin, and Chris Xu. "Multi-color femtosecond source for simultaneous excitation of multiple fluorescent proteins in two-photon fluorescence microscopy." In SPIE BiOS, edited by Ammasi Periasamy, Karsten König, and Peter T. C. So. SPIE, 2013. http://dx.doi.org/10.1117/12.2000583.

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Betke, Alexander, Bernd Voigt, Heiko Lokstein, and Ralf Menzel. "Two-photon fluorescence excitation spectroscopy of photosynthetic pigments and pigment-protein complexes." In 12th European Quantum Electronics Conference CLEO EUROPE/EQEC. IEEE, 2011. http://dx.doi.org/10.1109/cleoe.2011.5943251.

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Kawanoa, Hiroyuki, Yasuo Nabekawa, Akira Suda, Yu Oishia, Hideaki Mizuno, Atsushi Miyawaki, and Katsumi Midorikawa. "Adaptive control of two-photon fluorescence from green fluorescent protein by shaped femtosecond excitation pulses." In European Conference on Biomedical Optics. Washington, D.C.: OSA, 2003. http://dx.doi.org/10.1364/ecbo.2003.5143_10.

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Kawano, Hiroyuki, Yasuo Nabekawa, Akira Suda, Yu Oishi, Hideaki Mizuno, Atsushi Miyawaki, and Katsumi Midorikawa. "Adaptive control of two-photon fluorescence from green fluorescent protein by shaped femtosecond excitation pulses." In European Conference on Biomedical Optics 2003, edited by Albert-Claude Boccara. SPIE, 2003. http://dx.doi.org/10.1117/12.500445.

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Tanaka, M., J. Tada, F. Kannari, H. Kawano, H. Mizuno, A. Miyawaki, A. Suda, and K. Midorikawa. "Selective excitation of fluorescent proteins on the basis of the two-photon absorption spectrum measurement." In Frontiers in Optics. Washington, D.C.: OSA, 2006. http://dx.doi.org/10.1364/fio.2006.jwd52.

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Uegaki, Kumiko, Masahito Yamanaka, Kenta Saito, Nicholas I. Smith, Yoshiyuki Arai, Satoshi Kawata, Takeharu Nagai, and Katsumasa Fujita. "Simultaneous single and two-photon excitation of fluorescent proteins for multicolor imaging of cellular structures." In JSAP-OSA Joint Symposia. Washington, D.C.: OSA, 2014. http://dx.doi.org/10.1364/jsap.2014.18a_c4_10.

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Lakowicz, Joseph R., Ignacy Gryczynski, Henryk M. Malak, and Zygmunt Gryczynski. "Two-color two-photon excitation of fluorescence." In BiOS '97, Part of Photonics West, edited by Richard B. Thompson. SPIE, 1997. http://dx.doi.org/10.1117/12.273539.

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Yang, Mu-Han, Christopher G. L. Ferri, Payam A. Saisan, Maxim Abashin, Peifang Tian, Yeshayahu Fainman, and Anna Devor. "Multicolor two-photon excitation for increasing fluorescence excitation depth." In Optics and the Brain. Washington, D.C.: OSA, 2017. http://dx.doi.org/10.1364/brain.2017.brm4b.6.

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Liu, Yafeng, Tongsheng Chen, and Qingming Luo. "Fluorescence correlation spectroscopy based upon two-photon excitation." In Biomolecular photonoics and Multidimensional Microscopy, edited by Qingming Luo and Min Gu. SPIE, 2003. http://dx.doi.org/10.1117/12.546244.

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Meshalkin, Yuri P., E. E. Alfimov, D. E. Groshev, and V. K. Makukha. "Two-photon fluorescence excitation spectroscopy of biological molecules." In International Conference on Coherent and Nonlinear Optics, edited by Victor N. Zadkov. SPIE, 1996. http://dx.doi.org/10.1117/12.242166.

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Reports on the topic "Two-photon excitation fluorescence of proteins"

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DeArmond, Fredrick. Two-Photon Excitation, Fluorescence Microscopy, and Quantitative Measurement of Two-Photon Absorption Cross Sections. Portland State University Library, January 2000. http://dx.doi.org/10.15760/etd.5920.

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