TY - GEN
A1 - Engelhard, Sonja
A1 - Kumke, Michael Uwe
A1 - Löhmannsröben, Hans-Gerd
T1 - OPQS – optical process and quality sensing : exemplary applications in the beerbrewing and polyurethane foaming processes
N2 - Optical methods play an important role in process analytical technologies (PAT). Four examples of optical process and quality sensing (OPQS) are presented, which are based on three important experimental techniques: near-infrared absorption, luminescence quenching, and a novel method, photon density wave (PDW) spectroscopy. These are used to evaluate four process and quality parameters related to beer brewing and polyurethane (PU) foaming processes: the ethanol content and the oxygen (O2) content in beer, the biomass in a bioreactor, and the cellular structures of PU foam produced in a pilot production plant.
T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - paper 004
KW - process analytical technology
KW - beer
KW - biomass
KW - foam analysis
KW - NIR spectroscopy
KW - fluorescence quenching
KW - photon density wave spectroscopy
Y1 - 2006
U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-12191
ER -
TY - GEN
A1 - Schmälzlin, Elmar
A1 - Walz, Bernd
A1 - Klimant, Ingo
A1 - Schewe, Bettina
A1 - Löhmannsröben, Hans-Gerd
T1 - Monitoring hormone-induced oxygen consumption in the salivary glands of the blowfly, Calliphora vicina, by use of luminescent microbeads
N2 - The salivary glands of the blowfly were injected with luminescent oxygen-sensitive microbeads. The changes in oxygen content within individual gland tubules during hormone-induced secretory activity were quantified. The measurements are based on an upgraded phase-modulation technique, where the phase shift of the sensor phosphorescence is determined independently from concentration and background signals. We show that the combination of a lock-in amplifier with a fluorescence microscope results in a convenient setup to measure oxygen concentrations within living animal tissues at the cellular level.
T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - paper 006
Y1 - 2006
U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-12206
ER -
TY - GEN
A1 - Hildebrandt, Nico
A1 - Charbonnière, Loïc J.
A1 - Ziessel, Raymond F.
A1 - Löhmannsröben, Hans-Gerd
T1 - Quantum dots as resonance energy transfer acceptors for monitoring biological interactions
N2 - Quantum dots (QDs) are common as luminescing markers for imaging in biological applications because their optical properties seem to be inert against their surrounding solvent. This, together with broad and strong absorption bands and intense, sharp tuneable luminescence bands, makes them interesting candidates for methods utilizing Forster Resonance Energy Transfer (FRET), e. g. for sensitive homogeneous fluoroimmunoassays (FIA). In this work we demonstrate energy transfer from Eu3+-trisbipyridin (Eu-TBP) donors to CdSe-ZnS-QD acceptors in solutions with and without serum. The QDs are commercially available CdSe-ZnS core-shell particles emitting at 655 nm (QD655). The FRET system was achieved by the binding of the streptavidin conjugated donors with the biotin conjugated acceptors. After excitation of Eu-TBP and as result of the energy transfer, the luminescence of the QD655 acceptors also showed lengthened decay times like the donors. The energy transfer efficiency, as calculated from the decay times of the bound and the unbound components, amounted to 37%. The Forster-radius, estimated from the absorption and emission bands, was ca. 77Å. The effective binding ratio, which not only depends on the ratio of binding pairs but also on unspecific binding, was obtained from the donor emission dependent on the concentration. As serum promotes unspecific binding, the overall FRET efficiency of the assay was reduced. We conclude that QDs are good substitutes for acceptors in FRET if combined with slow decay donors like Europium. The investigation of the influence of the serum provides guidance towards improving binding properties of QD assays.
T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - paper 012
Y1 - 2006
U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-12213
ER -
TY - GEN
A1 - Schuler, Benjamin
A1 - Lipman, Everett A.
A1 - Steinbach, Peter J.
A1 - Kumke, Michael Uwe
A1 - Eaton, William A.
T1 - Polyproline and the "spectroscopic ruler" revisited with single-molecule fluorescence
N2 - To determine whether Förster resonance energy transfer (FRET) measurements can provide quantitative distance information in single-molecule fluorescence experiments on polypeptides, we measured FRET efficiency distributions for donor and acceptor dyes attached to the ends of freely diffusing polyproline molecules of various lengths. The observed mean FRET efficiencies agree with those determined from ensemble lifetime measurements but differ considerably from the values expected from Förster theory, with polyproline treated as a rigid rod. At donor–acceptor distances much less than the Förster radius R0, the observed efficiencies are lower than predicted, whereas at distances comparable to and greater than R0, they are much higher. Two possible contributions to the former are incomplete orientational averaging during the donor lifetime and, because of the large size of the dyes, breakdown of the point-dipole approximation assumed in Förster theory. End-to-end distance distributions and correlation times obtained from Langevin molecular dynamics simulations suggest that the differences for the longer polyproline peptides can be explained by chain bending, which considerably shortens the donor–acceptor distances.
T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - paper 008
KW - Förster resonance energy transfer
KW - molecular dynamics
KW - polypeptide
KW - FRET
Y1 - 2005
U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-12229
ER -
TY - GEN
A1 - Schmälzlin, Elmar
A1 - Dongen, Joost T. van
A1 - Klimant, Ingo
A1 - Marmodée, Bettina
A1 - Steup, Martin
A1 - Fishahn, Joachim
A1 - Geigenberger, Peter
A1 - Löhmannsröben, Hans-Gerd
T1 - An optical multifrequency phase-modulation method using microbeads for measuring intracellular oxygen concentrations in plants
N2 - A technique has been developed to measure absolute intracellular oxygen concentrations in green plants. Oxygen-sensitive phosphorescent microbeads were injected into the cells and an optical multifrequency phase-modulation technique was used to discriminate the sensor signal from the strong autofluorescence of the plant tissue. The method was established using photosynthesis-competent cells of the giant algae Chara corallina L., and was validated by application to various cell types of other plant species.
T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - paper 007
Y1 - 2005
U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-12232
ER -
TY - GEN
A1 - Lemke, Matthias
A1 - Fernández-Trujillo, Rebeca
A1 - Löhmannsröben, Hans-Gerd
T1 - In-situ LIF analysis of biological and petroleum-based hydraulic oils on soil
N2 - Absorption and fluorescence properties of 4 hydraulic oils (3 biological and 1 petroleum-based) were investigated. In-situ LIF (laser-induced fluorescence) analysis of the oils on a brown sandy loam soil was performed. With calibration, quantitative detection was achieved. Estimated limits of detection were below ca. 500 mg/kg for the petroleum-based oil and ca. 2000 mg/kg for one biological oil. A semi-quantitative classification scheme is proposed for monitoring of the biological oils. This approach was applied to investigate the migration of a biological oil in soil-containing compartments, namely a soil column and a soil bed.
T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - paper 011
KW - in-situ
KW - optical oil sensor
KW - lubricants
KW - hydraulic oils
KW - soil
KW - laser induced
Y1 - 2005
U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-12268
ER -
TY - GEN
A1 - Billard, Isabelle
A1 - Ansoborlo, Eric
A1 - Apperson, Kathleen
A1 - Arpigny, Sylvie
A1 - Azenha, M. Emilia
A1 - Birch, David
A1 - Bros, Pascal
A1 - Burrows, Hugh D.
A1 - Choppin, Gregory
A1 - Kumke, Michael Uwe
T1 - Aqueous solutions of Uranium(VI) as studied by time-resolved emission spectroscopy : a Round-Robin Test
N2 - Results of an inter-laboratory round-robin study of the application of time-resolved emission spectroscopy (TRES) to the speciation of uranium(VI) in aqueous media are presented. The round-robin study involved 13 independent laboratories, using various instrumentation and data analysis methods. Samples were prepared based on appropriate speciation diagrams and, in general, were found to be chemically stable for at least six months. Four different types of aqueous uranyl solutions were studied: (1) acidic medium where UO22+aq is the single emitting species, (2) uranyl in the presence of fluoride ions, (3) uranyl in the presence of sulfate ions, and (4) uranyl in aqueous solutions at different pH, promoting the formation of hydrolyzed species. Results between the laboratories are compared in terms of the number of decay components, luminescence lifetimes, and spectral band positions. The successes and limitations of TRES in uranyl analysis and speciation in aqueous solutions are discussed.
T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - paper 005
KW - uranium (VI)
KW - intercomparison
KW - speciation
Y1 - 2003
U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-12318
ER -
TY - GEN
A1 - Kumke, Michael Uwe
A1 - Frimmel, Fritz Hartmann
T1 - Stationary and time-resolved fluorescence for humic substances characterization
N2 - Steady-state and time-resolved fluorescence methods were applied to investigate the fluorescence properties of humic substances of different origins. Using standard 2D emission and total luminescence spectra, fluorescence maxima, the width of the fluorescence band and a relative fluorescence quantum efficiency were determined. Different trends for fulvic acids and humic acids were observed indicating differences in the heterogeneity of the sample fractions. The complexity of the fluorescence decay of humic substances is discussed and compared to simple model compounds. The effect of oxidation of humic substances on their fluorescence properties is discussed as well.
T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - paper 014
Y1 - 2002
U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-12353
ER -
TY - GEN
A1 - Dosche, Carsten
A1 - Löhmannsröben, Hans-Gerd
A1 - Bieser, A.
A1 - Dosa, P. I.
A1 - Han, S.
A1 - Iwamoto, M.
A1 - Schleifenbaum, A.
A1 - Vollhardt, K. Peter C.
T1 - Photophysical properties of [N]phenylenes
N2 - In the present study, photophysical properties of [N]phenylenes were studied by means of stationary and time-resolved absorption and fluorescence spectroscopy (in THF at room temperature). For biphenylene (1) and linear [3]phenylene (2a), internal conversion (IC) with quantum yields ΦIC > 0.99 is by far the dominant mechanism of S1 state deactivation. Angular [3]phenylene (3a), the zig-zag [4]- and [5]phenylenes (3b), (3c), and the triangular [4]phenylene (4) show fluorescence emission with fluorescence quantum yieds and lifetimes between ΦF = 0.07 for (3a) and 0.21 for (3c) and τF = 20 ns for (3a) and 81 ns for (4). Also, compounds (3) and (4) exhibit triplet formation upon photoexcitation with quantum yields as high as ΦISC = 0.45 for (3c). The strong differences in the fluorescence properties and in the triplet fromation efficiencies between (1) and (2a) on one hand and (3) and (4) on the other are related to the remarkable variation of the internal conversion (IC) rate constants kIC. A tentative classification of (1) and (2a) as “fast IC compounds”, with kIC > 109 s-1, and of (3) and (4) as “slow IC compounds”, with kIC ≈ 107 s-1, is suggested. This classification cannot simply be related to Hückel’s rule-type concepts of aromaticity, because the group of “fast IC compounds” consists of “antiaromatic” (1) and “aromatic” (2a), and the group of “slow IC compounds” consists of “antiaromatic” (3b), (4) and “aromatic” (3a), (3c). The IC in the [N]phenylenes is discussed within the framework of the so-called energy gap law established for non-radiative processes in benzenoid hydrocarbons.
T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - paper 001
Y1 - 2002
U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-11936
ER -
TY - GEN
A1 - Löhmannsröben, Hans-Gerd
A1 - Beitz, Toralf
A1 - Luadien, Robert
A1 - Schultze, Rainer
T1 - Laser-based ion mobility spectrometry for sensing of aromatic compounds
N2 - The drift time spectra of polycyclic aromatic hydrocarbons (PAH), alkylbenzenes and alkylphenylethers were recorded with a laser-based ion mobility (IM) spectrometer. The ion mobilities of all compounds were determined in helium as drift gas. This allows the calculation of the diffusion cross sections (Omegacalc) on the basis of the exact hard sphere scattering model (EHSSM) and their comparison with the experimentally determined diffusion cross sections (Omegaexp). These Omegaexp/Omegacalc-correlations are presented for molecules with a rigid structure like PAH and prove the reliability of the theoretical model and experimental method. The increase of the selectivity of IM spectrometry is demonstrated using resonance enhanced multiphoton ionisation (REMPI) at atmospheric pressure, realized by tuneable lasers. The REMPI spectra of nine alkylbenzenes and alkylphenylethers are investigated. On the basis of these spectra, the complete qualitative distinction of eight compounds in a mixture is shown. These experiments are extended to alkylbenzene isomer mixtures.
T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - paper 003
KW - Laser
KW - REMPI
KW - Ion mobility spectrometry
KW - PAH
Y1 - 2004
U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-11892
ER -
TY - GEN
A1 - Dosche, Carsten
A1 - Kumke, Michael Uwe
A1 - Löhmannsröben, Hans-Gerd
A1 - Ariese, F.
A1 - Bader, Arjen N.
A1 - Gooijer, C.
A1 - Miljanic, O. S.
A1 - Iwamoto, M.
A1 - Vollhardt, K. Peter C.
A1 - Puchta, R. ; van Eikema Hommes, N. J. R.
T1 - Deuteration effects on the vibronic structure of the fluorescence spectra and the internal conversion rates of triangular [4]Phenylene
N2 - Deuteration effects on the vibronic structure of the emission and excitation spectra of triangular [4]phenylene (D3h[4]phenylene) were studied using laser-excited Shpolskii spectroscopy (LESS) in an octane matrix at 4.2 K. For correct assignment of the vibrational modes, the experimental results were compared with calculated frequencies (B3LYP/6-31G*). CH vibrations were identified by their characteristic isotopic shifts in the spectra of deuterated triangular [4]phenylenes. Two CC stretching modes, at 100 cm–1 and 1176 cm–1, suitable as probes for bond strength changes in the excited state, were identified. The isotope effect on the internal conversion rates of triangular [4]phenylene was evaluated from measurements of temperature dependent lifetime. Isotope dependency and the magnitude of the internal conversion rates indicate that internal conversion in triangular [4]phenylene is most likely induced by CH vibrations. The results obtained by LESS and lifetime measurements were compared with PM3 PECI calculations of the excited state structure. The theoretical results and the relation between ground and excited state vibration energies of the 1176 cm–1 probe vibration indicate a reduction of bond alternation of the central cyclohexatriene ring in the excited state.
T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - paper 002
Y1 - 2004
U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-11881
ER -
TY - GEN
A1 - Hörner, Gerald
A1 - Lau, Steffen
A1 - Kantor, Zoltan
A1 - Löhmannsröben, Hans-Gerd
T1 - Isotope selective analysis of CO2 with tunable diode laser (TDL) spectroscopy in the NIR
N2 - The performance of a home-built tunable diode laser (TDL) spectrometer, aimed at multi-line detection of carbon dioxide, has been evaluated and optimized. In the regime of the (3001)III / (000) band of 12CO2 around 1.6 μm, the dominating isotope species 12CO2, 13CO2, and 12C18O16O were detected simultaneously without interference by water vapor. Detection limits in the range of few ppmv were obtained for each species utilizing wavelength modulation (WM) spectroscopy with balanced detection in a long-path absorption cell set-up. High sensitivity in conjunction with high precision —typically ±1‰ and ±6‰ for 3% and 0.7% of CO2, respectively— renders this experimental approach a promising analytical concept for isotope-ratio determination of carbon dioxide in soil and breath gas. For a moderate 12CO2 line, the pressure dependence of the line profile was characterized in detail, to account for pressure effects on sensitive measurements.
T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - paper 020
KW - Isotopenverhältnis
KW - Kohlendioxid
KW - Diodenlaserspektroskopie
KW - Isotope ratio
KW - carbon monoxide
KW - diode laser spectroscopy
Y1 - 2004
U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-10167
ER -
TY - GEN
A1 - Niederkrüger, Matthias
A1 - Salb, Christian
A1 - Beck, Michael
A1 - Hildebrandt, Niko
A1 - Löhmannsröben, Hans-Gerd
A1 - Marowsky, Gerd
T1 - Improvement of a fluorescence immunoassay with a compact diode-pumped solid state laser at 315 nm
N2 - We demonstrate the improvement of fluorescence immunoassay (FIA) diagnostics in deploying a newly developed compact diode-pumped solid state (DPSS) laser with emission at 315 nm. The laser is based on the quasi-three-level transition in Nd:YAG at 946 nm. The pulsed operation is either realized by an active Q-switch using an electro-optical device or by introduction of a Cr4+:YAG saturable absorber as passive Q-switch element. By extra-cavity second harmonic generation in different nonlinear crystal media we obtained blue light at 473 nm. Subsequent mixing of the fundamental and the second harmonic in a β-barium-borate crystal provided pulsed emission at 315 nm with up to 20 μJ maximum pulse energy and 17 ns pulse duration. Substitution of a nitrogen laser in a FIA diagnostics system by the DPSS laser succeeded in considerable improvement of the detection limit. Despite significantly lower pulse energies (7 μJ DPSS laser versus 150 μJ nitrogen laser), in preliminary investigations the limit of detection was reduced by a factor of three for a typical FIA.
T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - paper 016
KW - Immunoassay
KW - Fluoreszenz-Resonanz-Energie-Transfer
KW - Neodym-YAG-Laser
KW - 946 nm
KW - 473 nm
KW - 315 nm
KW - gepulster DPSS Laser
KW - sättigbarer Absorber
KW - fluorescence immunoassay
KW - 946 nm
KW - 473 nm
KW - 315 nm
KW - pulsed DPSS laser
Y1 - 2006
U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-10150
ER -
TY - GEN
A1 - Hörner, Gerald
A1 - Lau, Steffen
A1 - Löhmannsröben, Hans-Gerd
T1 - NIR-diode laser spectroscopy for isotope-selective sensing of soil-respired carbon dioxide
N2 - The performance of a home-built tunable diode laser (TDL) spectrometer has been optimized regarding multi-line detection of carbon dioxide in natural gases. In the regime of the (3001)III ← (000) band of 12CO2 around 1.6 μm, the dominating isotope species 12CO2, 13CO2, and 12C18O16O were detected simultaneously. In contrast to most established techniques, selective measurements are performed without any sample preparation. This is possible since the CO2 detection is free of interference from water, ubiquitous in natural gases. Detection limits in the range of a few ppmv were obtained for each species utilizing wavelength modulation (WM) spectroscopy with balanced detection in a long-path absorption cell set-up. Linear calibration plots cover a dynamic range of four orders of magnitude, allowing for quantitative CO2 detection in various samples, like soil and breath gas. High isotopic resolution enables the excellent selectivity, sensitivity, and stability of the chosen analytical concept. The obtained isotopic resolution of typically ± 1.0 ‰ and ± 1.5 ‰ (for 3 vol. % and 0.7 vol. % of CO2, respectively) offers a promising analytical tool for isotope-ratio determination of carbon dioxide in soil gas. Preliminary experiments on soil respiration for the first time combine the on-line quantification of the overall carbon dioxide content with an optode sensor and isotopic determination (TDL system) of natural gas species.
T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - paper 021
KW - Kohlendioxid
KW - Isotopenverhältnis
KW - Bodengas
KW - Diodenlaserspektroskopie
KW - tunable diode laser (TDL)
KW - carbon dioxide
KW - isotope ratios
KW - soil gas
Y1 - 2004
U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-10148
ER -
TY - GEN
A1 - Lau, Steffen
A1 - Salffner, Katharina
A1 - Löhmannsröben, Hans-Gerd
T1 - Isotopic resolution of carbon monoxide and carbon dioxide by NIR diode laser spectroscopy
N2 - Near-infrared (NIR) absorption spectroscopy with tunable diode lasers allows the simultaneous detection of the three most important isotopologues of carbon dioxide (12CO2, 13CO2, 12C18O16O) and carbon monoxide (12CO, 13CO, 12C18O). The flexible and compact fiber-optic tunable diode laser absorption spectrometer (TDLAS) allows selective measurements of CO2 and CO with high isotopic resolution without sample preparation since there is no interference with water vapour. For each species, linear calibration plots with a dynamic range of four orders of magnitude and detection limits (LOD) in the range of a few ppm were obtained utilizing wavelength modulation spectroscopy (WMS) with balanced detection in a Herriott-type multipass cell. The high performance of the apparatus is illustrated by fill-evacuation-refill cycles.
T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - paper 017
KW - Isotop
KW - Kohlenmonoxid
KW - Kohlendioxid
KW - Diodenlaserspektroskopie
KW - isotope
KW - carbon monoxide
KW - carbon dioxide
KW - diode laser spectroscopy
Y1 - 2006
U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-10135
ER -
TY - GEN
A1 - Löhmannsröben, Hans-Gerd
A1 - Beck, Michael
A1 - Hildebrandt, Niko
A1 - Schmälzlin, Elmar
A1 - van Dongen, Joost T.
T1 - New challenges in biophotonics : laser-based fluoroimmuno analysis and in-vivo optical oxygen monitoring
N2 - Two examples of our biophotonic research utilizing nanoparticles are presented, namely laser-based fluoroimmuno analysis and in-vivo optical oxygen monitoring. Results of the work include significantly enhanced sensitivity of a homogeneous fluorescence immunoassay and markedly improved spatial resolution of oxygen gradients in root nodules of a legume species.
T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - paper 018
KW - Sauerstoff
KW - Quantenpunkt
KW - Lumineszenz
KW - Immunoassay
KW - Energietransfer
KW - Fluoreszenz-Resonanz-Energie-Transfer
KW - Nanopartikel
KW - Lanthanoide
KW - Optode
KW - Förster Resonanz Energie Transfer
KW - Biophotonik
KW - biophotonics
KW - nanoparticles
KW - immunoassay
KW - oxygen
KW - optode
Y1 - 2006
U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-10120
ER -
TY - GEN
A1 - Beck, Michael
A1 - Hildebrandt, Niko
A1 - Löhmannsröben, Hans-Gerd
T1 - Quantum dots as acceptors in FRET-assays containing serum
N2 - Quantum dots (QDs) are common as luminescing markers for imaging in biological applications because their optical properties seem to be inert against their surrounding solvent. This, together with broad and strong absorption bands and intense, sharp tuneable luminescence bands, makes them interesting candidates for methods utilizing Förster Resonance Energy Transfer (FRET), e. g. for sensitive homogeneous fluoroimmunoassays (FIA). In this work we demonstrate energy transfer from Eu3+-trisbipyridin (Eu-TBP) donors to CdSe-ZnS-QD acceptors in solutions with and without serum. The QDs are commercially available CdSe-ZnS core-shell particles emitting at 655 nm (QD655). The FRET system was achieved by the binding of the streptavidin conjugated donors with the biotin conjugated acceptors. After excitation of Eu-TBP and as result of the energy transfer, the luminescence of the QD655 acceptors also showed lengthened decay times like the donors. The energy transfer efficiency, as calculated from the decay times of the bound and the unbound components, amounted to 37%. The Förster-radius, estimated from the absorption and emission bands, was ca. 77 Å. The effective binding ratio, which not only depends on the ratio of binding pairs but also on unspecific binding, was obtained from the donor emission dependent on the concentration. As serum promotes unspecific binding, the overall FRET efficiency of the assay was reduced. We conclude that QDs are good substitutes for acceptors in FRET if combined with slow decay donors like Europium. The investigation of the influence of the serum provides guidance towards improving binding properties of QD assays.
T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - paper 019
KW - Quantenpunkt
KW - Lumineszenz
KW - Serum
KW - Europium
KW - Immunoassay
KW - Energietransfer
KW - Fluoreszenz-Resonanz-Energie-Transfer
KW - Förster-Resonanz-Energie-Transfer
KW - Quantum Dot
KW - Luminescence
KW - Serum
KW - Europium
KW - Immunoassay
KW - Energy Transfer
KW - FRET
Y1 - 2006
U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-9504
ER -
TY - THES
A1 - Maltseva, Elena
T1 - Model membrane interactions with ions and peptides at the air/water interface
T1 - Wechselwirkungen von Modellmembranen mit Ionen und Peptiden - studiert an der Luft-Wasser-Grenzfläche
N2 - The interactions between peptides and lipids are of fundamental importance in the functioning of numerous membrane-mediated biochemical processes including antimicrobial peptide action, hormone-receptor interactions, drug bioavailability across the blood-brain barrier and viral fusion processes. Alteration of peptide structure could be a cause of many diseases. Biological membranes are complex systems, therefore simplified models may be introduced in order to understand processes occurring in nature. The lipid monolayers at the air/water interface are suitable model systems to mimic biological membranes since many parameters can be easily controlled. In the present work the lipid monolayers were used as a model membrane and their interactions with two different peptides B18 and Amyloid beta (1-40) peptide were investigated. B18 is a synthetic peptide that binds to lipid membranes that leads to the membrane fusion. It was demonstrated that it adopts different structures in the aqueous solutions and in the membrane interior. It is unstructured in solutions and forms alpha-helix at the air/water interface or in the membrane bound state. The peptide has affinity to the negatively charged lipids and even can fold into beta-sheet structure in the vicinity of charged membranes at high peptide to lipid ratio. It was elucidated that in the absence of electrostatic interactions B18 does not influence on the lipid structure, whereas it provides partial liquidization of the negatively charged lipids. The understanding of mechanism of the peptide action in model system may help to develop the new type of antimicrobial peptides as well as it can shed light on the general mechanisms of peptide/membrane binding. The other studied peptide - Amyloid beta (1-40) peptide, which is the major component of amyloid plaques found in the brain of patients with Alzheimer's disease. Normally the peptide is soluble and is not toxic. During aging or as a result of the disease it aggregates and shows a pronounced neurotoxicity. The peptide aggregation involves the conformational transition from a random coil or alpha-helix to beta-sheets. Recently it was demonstrated that the membrane can play a crucial role for the peptide aggregation and even more the peptide can cause the change in the cell membranes that leads to a neuron death. In the present studies the structure of the membrane bound Amyloid beta peptide was elucidated. It was found that the peptide adopts the beta-sheet structure at the air/water interface or being adsorbed on lipid monolayers, while it can form alpha-helical structure in the presence of the negatively charged vesicles. The difference between the monolayer system and the bulk system with vesicles is the peptide to lipid ratio. The peptide adopts the helical structure at low peptide to lipid ratio and folds into beta-sheet at high ratio. Apparently, Abeta peptide accumulation in the brain is concentration driven. Increasing concentration leads to a change in the lipid to peptide ratio that induces the beta-sheet formation. The negatively charged lipids can act as seeds in the plaque formation, the peptide accumulates on the membrane and when the peptide to lipid ratio increases it the peptide forms toxic beta-sheet containing aggregates.
N2 - Wechselwirkungen zwischen Peptiden und Lipiden sind von grundlegender Bedeutung für die Funktion vieler Membran-vermittelter biochemischer Prozesse wie der Wirkung von antimikrobiellen Peptiden, Hormon-Rezeptor Wechselwirkungen, Bioverfügbarkeit von Arzneistoffen durch die Blut-Hirn-Schranke und viraler Fusionsprozesse. Veränderungen in der Peptidstruktur können die Ursache für viele Erkrankungen sein. Biologische Membranen sind für grundlegende physikalisch-chemische Untersuchungen von Naturprozessen zu komplexe Systeme, so dass vereinfachte Modelle für solche Studien eingesetzt werden. Eine Lipidmonoschicht an der Wasser/Luft Grenzfläche ist ein geeignetes Modellsystem für eine Membranoberfläche. Viele physikalisch-chemischen Parameter können auf einfache Weise gezielt verändert werden. In der vorliegenden Arbeit wurden Lipidmonoschichten genutzt, um Wechselwirkungen mit zwei unterschiedlichen Peptiden (B18 and Amyloid Beta (1-40) Peptid) zu untersuchen. B18 ist ein oberflächenaktives synthetisches Peptid, das an Lipidmembranen bindet und zu Membranfusion führt. Es kann verschiedene Sekundärstrukturen ausbilden. So ist B18 in wässrigen Lösungen ungeordnet und bildet eine alpha-helikale Struktur an der Wasser/Luft Grenzfläche. Das Peptid hat eine große Affinität zu negativ geladenen Lipiden und kann in der Nähe von geladenen Membranoberflächen bei einem großen Peptid/Lipid Verhältnis eine Beta-Faltblatt Struktur ausbilden. Beim Fehlen elektrostatischer Wechselwirkungen hat B18 keinen Einfluss auf die Lipidstruktur. Es wirkt jedoch strukturabbauend auf anionische Lipide. Das Verständnis der Peptidwirkungen in Modellsystemen kann helfen, generelle Mechanismen von Peptide-Membran Wechselwirkungen zu verstehen und zur Entwicklung neuer antimikrobieller Peptide beizutragen. Amyloid Beta (1-40) Peptid ist die Hauptkomponente von Amyloid-Plaque, das im Gehirn von Alzheimer Patienten gefunden wird. Normalerweise ist das Peptid löslich und nicht toxisch. Hohe Neurotoxizität wird bei Peptidaggregation, die eine Strukturumwandlung von ungeordnet oder alpha-helikal zu Beta-Faltblatt nach sich zieht, beobachtet. In der vorliegenden Arbeit wurde die Struktur des Membran-gebundenen Amyloid Beta (1-40) Peptids untersucht. Es zeigte sich, dass das Peptid nach Adsorption an die Wasser/Luft Grenzfläche oder an Lipidmonoschichten eine Beta-Faltblatt Struktur ausbildet. Eine alpha-helikale Sekundärstruktur wird nur bei Anwesenheit negativ geladenen Lipidvesikel gefunden. Der entscheidende Unterschied zwischen den Monoschicht- und Vesikel-Systemen ist das Peptid/Lipid Verhältnis. Die alpha-helikale Struktur wird nur bei kleinem Peptid/Lipid Verhältnis beobachtet, während bei großem eine Beta-Faltblatt Struktur auftritt. Steigende Konzentration an Amyloid Beta (1-40) Peptid führt zum Anstieg des Peptid/Lipid Verhältnisses und damit zur Ausbildung der Beta-Faltblatt Struktur. Negativ geladene Lipide können somit als Keimpunkte für die Plaquebildung fungieren.
KW - Lipide
KW - Monoschicht
KW - Peptide
KW - Phospholipid
KW - Langmuir monolayers
KW - IRRAS
KW - Amyloid peptide
Y1 - 2005
U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus-5670
ER -