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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 (30<SUP>0</SUP>1)<SUB>III</SUB> ← (000) band of <SUP>12</SUP>CO<SUB>2</SUB> around 1.6 μm, the dominating isotope species <SUP>12</SUP>CO<SUB>2</SUB>, <SUP>13</SUP>CO<SUB>2</SUB>, and <SUP>12</SUP>C<SUP>18</SUP>O<SUP>16</SUP>O were detected simultaneously. In contrast to most established techniques, selective measurements are performed without any sample preparation. This is possible since the CO<SUB>2</SUB> 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 CO<SUB>2</SUB> 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 CO<SUB>2</SUB>, 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.
Near-infrared (NIR) absorption spectroscopy with tunable diode lasers allows the simultaneous detection of the three most important isotopologues of carbon dioxide (<SUP>12</SUP>CO<SUB>2</SUB>, <SUP>13</SUP>CO<SUB>2</SUB>, <SUP>12</SUP>C<SUP>18</SUP>O<SUP>16</SUP>O) and carbon monoxide (<SUP>12</SUP>CO, <SUP>13</SUP>CO, <SUP>12</SUP>C<SUP>18</SUP>O). The flexible and compact fiber-optic tunable diode laser absorption spectrometer (TDLAS) allows selective measurements of CO<SUB>2</SUB> 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.
Löhmannsröben, Hans-Gerd ; Beck, Michael ; Hildebrandt, Niko ; Schmälzlin, Elmar ; van Dongen, Joost T.
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.
Nonaqueous synthesis of metal oxide nanoparticles and their assembly into mesoporous materials
(2006)
This thesis mainly consist of two parts, the synthesis of several kinds of technologically interesting crystalline metal oxide nanoparticles via nonaqueous sol-gel process and the formation of mesoporous metal oxides using some of these nanoparticles as building blocks via evaporation induced self-assembly (EISA) technique. In the first part, the experimental procedures and characterization results of successful syntheses of crystalline tin oxide and tin doped indium oxide (ITO) nanoparticles are reported. SnO2 nanoparticles exhibit monodisperse particle size (3.5 nm in average), high crystallinity and particularly high dispersibility in THF, which enable them to become the ideal particulate precursor for the formation of mesoporous SnO2. ITO nanoparticles possess uniform particle morphology, narrow particle size distribution (5-10 nm), high crystallinity as well as high electrical conductivity. The synthesis approaches and characterization of various mesoporous metal oxides, including TiO2, SnO2, mixture of CeO2 and TiO2, mixture of BaTiO3 and SnO2, are reported in the second part of this thesis. Mesoporous TiO2 and SnO2 are presented as highlights of this part. Mesoporous TiO2 was produced in the forms of both films and bulk material. In the case of mesoporous SnO2, the study was focused on the high order of the porous structure. All these mesoporous metal oxides show high crystallinity, high surface area and rather monodisperse pore sizes, which demonstrate the validity of EISA process and the usage of preformed crystalline nanoparticles as nanobuilding blocks (NBBs) to produce mesoporous metal oxides.
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 Eu<SUP>3+</SUP>-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.
In this work approaches for new detection system development for an Analytical Ultracentrifuge (AUC) were explored. Unlike its counterpart in chromatography fractionation techniques, the use of a Multidetection system for AUC has not yet been implemented to full extent despite its potential benefit. In this study we tried to couple existing fundamental spectroscopic and scattering techniques that are used in day to day science as tool for extracting analyte information. Trials were performed for adapting Raman, Light scattering and UV/Vis (with possibility to work with the whole range of wavelengths) to AUC. Conclusions were drawn for Raman and Light scattering to be a possible detection system for AUC, while the development for a fast fiber optics based multiwavelength detector was completed. The multiwavelength detector demonstrated the capability of data generation matching the literature and reference measurement data and faster data collection than that of the commercial instrument. It became obvious that with the generation of data in 3-D space in the UV/Vis detection system, the user can select the wavelength for the evaluation of experimental results as the data set contains the whole range of information from UV/Vis wavelength. The detector showed the data generation with much faster speed unlike the commercial instruments. The advantage of fast data generation was exemplified with the evaluation of data for a mixture of three colloids. These data were in conformity with measurement results from normal radial experiments and without significant diffusion broadening. Thus conclusions were drawn that with our designed Multiwavelength detector, meaningful data in 3-D space can be collected with much faster speed of data generation.
Retrosynthese von Perlmutt
(2006)
In dieser Arbeit ist es gelungen, die Bedeutung physikalisch-chemischer Mechanismen in der Biomineralisation gegenüber der bisher angenommenen Dominanz spezifischer biomolekularer Erkennungsmechanismen aufzuzeigen. Dazu wurden drei Ansätze verfolgt: Zum einen wurden Studien zur Calciumcarbonatkristallisation durchgeführt. Zum anderen wurde das Biomineral Perlmutt intensiv untersucht. Als drittes wurde ein Modellsystem entwickelt, mit dem künstliches Perlmutt synthetisiert und ein Mechanismus für die Perlmuttmineralisation vorgeschlagen werden konnte. Im ersten Schritt wurden in einem simplen Kristallisationsansatz komplexe Calciumcarbonatüberstrukturen ohne die Verwendung von Additiven synthetisiert. Es wurde gezeigt, daß diese durch orientierte Anlagerung von Nanopartikeln gebildet werden, bei der dipolare Felder eine wichtige Rolle zu spielen scheinen. Dieser Mechansimus war bislang für Calciumcarbonat unbekannt und ermöglicht die Synthese komplexer Kristallmorphologien, wodurch die Frage aufgeworfen wird, ob er bei der Biomineralbildung von Bedeutung sein kann. Durch Einsatz minimaler Mengen eines einfachen, synthetischen Additivs bei der Kristallisation wurden zu Überstrukturen angeordnete Aragonitplättchen synthetisiert, die von einer wenige nm dicken Schicht aus amorphen Calciumcarbonat umgeben sind. Eine solche Schicht wurde auch bei den Aragonitplättchen Perlmutts entdeckt (s.u.) und bietet möglicherweise in verschiedenen Systemen eine Erklärung für die Stabilisierung der sonst metastabilen Aragonitphase. Im zweiten Schritt wurden bei der Untersuchung von natürlichem Perlmutt zwei bislang unbekannte Strukturmerkmale entdeckt: Es gibt Bereiche, die nicht aus den charakteristischen Plättchen bestehen, sondern wesentlich weniger stark mineralisert sind. Die Mineralphase besteht in diesen Bereichen aus Nanopartikeln. Es wurde weiterhin gezeigt, daß die Aragonitplättchen von einer wenige nm dicken Schicht aus amorphem Calciumcarbonat umgeben ist. Die gängigen Modelle der Perlmuttbildung sind mit diesen Beobachtungen nicht zu vereinbaren und somit zu hinterfragen. Dagegen deuten diese Ergebnisse ein Wachstum von Perlmutt über ACC-Nanopartikel an. Unter der Annahme der Bedeutung physikalisch-chemischer Mechanismen in der Biomineralisation wurde schließlich als dritter Schritt ein Ansatz zur in vitro-Retrosynthese von Biomineralien ausgehend von ihrer unlöslichen Matrix entwickelt. Mit diesem Ansatz ist es erstmals gelungen, künstliches Perlmutt auf synthetischem Wege herzustellen, das morphologisch nicht vom Original zu unterscheiden ist. Die existierenden Unterschiede konnten zeigen, daß der Mineralisationsprozeß nicht auf ein spezifisches Mikroumgebungssystem beschränkt, sondern "allgemeiner gültig"' sein muß. Bei der Retrosynthese gibt es zwei Schlüsselfaktoren: Zum einen die demineralisierte unlösliche Perlmuttmatrix als dreidimensionales Gerüst für das künstliche Perlmutt, zum anderen amorphe Precursorpartikel, die die Mineralphase bilden. Es werden keinerlei Proteine oder andere Biomoleküle verwendet. Dieser Ansatz bietet die Möglichkeit, den Mineralisationsprozeß an einem in vitro-Modellsystem zu verfolgen, was für das in vivo-System, wenn überhaupt, nur unter starken Einschränkungen möglich ist. Es wurde gezeigt, daß das künstliche Perlmutt über die Mesoskalentransformation von ACC-Precursorn innerhalb der Matrix gebildet wird und als möglicher Mechanismus bei der Biomineralisation von natürlichem Perlmutt diskutiert. Es konnte in der vorliegenden Arbeit konsequent gezeigt werden, daß die Imitation von Biomineralisationsprozessen in in vitro-Ansätzen möglich ist, wobei chemisch-physikalische Parameter dominieren. In zukünftigen Studien sollten einerseits die mechanischen Eigenschaften des künstlichen Perlmutts untersucht werden, wofür sich in Vorversuchen im Rahmen dieser Arbeit die Nanoindentierung als geeignet herausgestellt hat. Es sollte geprüft werden, ob das hier ermittelte Prinzip zur Mineralisierung in der Materialentwicklung angewendet werden kann. Andererseits sollte die Retrosynthese auf andere Systeme ausgeweitet und in vivo-Studien durchgeführt werden, um die Gültigkeit der vorgeschlagenen Mechanismen zu überprüfen.
Ultrathin, semi-permeable membranes are not only essential in natural systems (membranes of cells or organelles) but they are also important for applications (separation, filtering) in miniaturized devices. Membranes, integrated as diffusion barriers or filters in micron scale devices need to fulfill equivalent requirements as the natural systems, in particular mechanical stability and functionality (e.g. permeability), while being only tens of nm in thickness to allow fast diffusion times. Promising candidates for such membranes are polyelectrolyte multilayers, which were found to be mechanically stable, and variable in functionality. In this thesis two concepts to integrate such membranes in larger scale structures were developed. The first is based on the directed adhesion of polyelectrolyte hollow microcapsules. As a result, arrays of capsules were created. These can be useful for combinatorial chemistry or sensing. This concept was expanded to couple encapsulated living cells to the surface. The second concept is the transfer of flat freestanding multilayer membranes to structured surfaces. We have developed a method that allows us to couple mm2 areas of defect free film with thicknesses down to 50 nm to structured surfaces and to avoid crumpling of the membrane. We could again use this technique to produce arrays of micron size. The freestanding membrane is a diffusion barrier for high molecular weight molecules, while small molecules can pass through the membrane and thus allows us to sense solution properties. We have shown also that osmotic pressures lead to membrane deflection. That could be described quantitatively.
Der Adhäsionsprozess an Cellulosefasern sollte bis hin zur molekularen Ebene aufgeklärt werden. Zuerst wurde eine definierte Celluloseoberfläche durch Aufschleudern hergestellt und eingehend charakterisiert wurde. Die Beschaffenheit der Oberfläche ist stark abhängig von der Konzentration Celluloselösung, die für das Aufschleudern benutzt wurde. Für geringe Konzentrationen bildete sich ein Netzwerk von Cellulosefibrillen über die Siliziumoberfläche aus. In einem mittleren Bereich wurden geschlossene Filme mit Dicken zwischen 1,5 und ca. 10 nm erhalten. Diese Filme wiesen eine Rauhigkeit unter 2 nm auf, was der Größe eines Cellulosemoleküls entspräche. Mit höheren Konzentrationen wurden die Filme dicker und auch rauer. Die molekular glatten Cellulosefilme wurden zur Untersuchung der Adhäsionseigenschaften zwischen Celluloseoberflächen und ihrer Modifikation durch den Einfluss von Haftvermittlern (Polyelektrolyte) eingesetzt. Dafür wurde das Haftungsvermögen von Celluloseperlen auf der glatten Celluloseoberfläche analysiert. Die Adhäsion der Celluloseperlen hängt dabei von der Menge und von der Art des adsorbierten Polyelektrolyts ab. Geringe Polyelektrolytkonzentrationen ermöglichen wegen der teilweisen Bedeckung der Celluloseoberflächen eine Verstärkung der Haftvermittlung (Adhäsion). Höhere Konzentrationen an Polyelektrolyten bedecken die Celluloseoberfläche komplett, so dass die Symmetrie der Polyelektrolytadsorption entscheidend wird. Wenn Celluloseperlen und glatte Celluloseoberfläche gleichzeitig mit Polyelektrolyten beschichtet werden verringert sich bei hohen Konzentrationen die Adhäsion erheblich. Ein hohes Maß an Adhäsion wird dagegen erreicht, wenn nur eine der beiden gegenüberliegende Celluloseoberflächen separat beschichtet und dann in Kontakt mit der anderen gebracht Oberfläche wurde. Der Einfluss einer äußeren Kraft auf die haftenden Celluloseperlen wurde untersucht, um eine Aussage über die auftretenden Adhäsionskräfte machen zu können. Dafür wurde eine spezielle Strömungszelle entwickelt, mit der die Bewegung von Celluloseperlen beobachtet werden kann. Je nach Ausrichtung der Strömungszelle bewegten sich die Celluloseperlen unterschiedlich schnell. War die Strömungszelle horizontal ausgerichtet, wirkte zusätzlich zur Adhäsionskraft noch die Gravitation auf die Oberfläche, so dass hier eine wesentlich langsamere Durchschnittsgeschwindigkeit ermittelt wurde als für die Messungen in vertikaler Ausrichtung. Die Zugabe von verschiedenen Polyelektrolyten bei den Messungen mit der Strömungszelle hatte einen erheblichen Einfluss auf die Beweglichkeit der haftenden Celluloseperlen. Bei geringen Polyelektrolytkonzentrationen wurde das Potential der einzelnen Polyelektrolyten zur effektiven Haftvermittlung durch eine signifikant geringere Durchschnittsgeschwindigkeit der rollenden Celluloseperlen sichtbar. Aus der kritischen Anfangsgeschwindigkeit der rollenden Celluloseperlen wurde die Adhäsionsarbeit errechnet, die für die Bewegung nötig ist.
Die Beeinflussung optischer Eigenschaften durch Bestrahlung stellt eine Grundlage für die Herstellung anisotroper optischer Komponenten dar. In dünnen Schichten von Azobenzen-Polymeren kann optische Anisotropie durch linear polarisierte Bestrahlung induziert oder modifiziert werden. Ziel der Arbeit war es, wesentliche Struktur-Eigenschafts-Beziehungen zum Prozess der Photoorientierung zu erarbeiten, um so eine Optimierung der Materialien für verschiedene Anwendungen ermöglichen. In isotropen Schichten flüssigkristalliner und amorpher Azobenzen-Polymeren wird das Ausmaß der induzierten optischen Anisotropie günstig durch eine Donor/Akzeptor-Substitution in 4,4'-Position beeinflusst. Die Induktionsgeschwindigkeit ist in Schichten flüssigkristalliner Polymeren deutlich geringer, jedoch lassen sich höhere Werte der Doppelbrechung und des Dichroismus erreichen. In Copolymeren bewirkt die Photoorientierung der Azobenzen-Seitengruppen eine kooperative Orientierung von formanisotropen Seitengruppen. Die Mesogenität der nicht-photochromen Seitengruppen erhöht das Ausmaß der induzierten optischen Anisotropie. Die Stabilität der induzierten Doppelbrechung und des Dichroismus wird durch diese Gruppen gesteigert. In Schichten flüssigkristalliner Polymeren wird die induzierte optische Anisotropie beim Tempern im Bereich der Mesophasen erheblich verstärkt. Dabei reicht ein geringes Maß an induzierter Anisotropie aus, um Doppelbrechungs- und Dichroismuswerte zu erzielen, wie sie für LC-Domänen typisch sind. In orientierten Schichten von Azobenzen-Polymeren wird das Resultat der linear polarisierten Bestrahlung durch die Stärke der anisotropen Wechselwirkungen in den flüssigkristallinen Domänen oder den LB-Multilayern bestimmt. Eine lichtinduzierte Reorientierung kann nur erreicht werden, wenn diese Wechselwirkungen überwunden werden können. Erfolgt eine Photoreorientierung in den orientierten Polymerschichten, werden in Copolymeren formanisotrope Seitengruppen ebenfalls kooperativ reorientiert. Eine vorgelagerte UV-Bestrahlung kann durch Erzeugung eines hohen Anteils an nicht-mesogenen Z-Isomeren die anisotropen Wechselwirkungen stark schwächen und so die Seitengruppen entkoppeln. Aus diesem Zustand erfolgt die Photoreorientierung mit einer der Photoorientierung in isotropen Schichten vergleichbaren Effizienz. Die erarbeiteten Struktur-Eigenschafts-Beziehungen liefern einen Beitrag zur Optimierung derartiger Materialien für Anwendungen in den Bereichen optischer Funktionsschichten, holographischer Datenspeicherung oder der Generierung von Oberflächenreliefgittern.