TY - THES A1 - Schulte, Franziska T1 - Raman-Spektroskopie als Werkzeug für die Charakterisierung und Klassifizierung von Pollen N2 - Diese Arbeit widmet sich der Analyse von Pollenkörnern und einzelnen Pollenkomponenten mit Hilfe der Raman-Spektroskopie, ergänzt wurden diese Untersuchungen durch den Einsatz der Massenspektrometrie, ESEM-Technik, Hochleistungs-Dünnschicht- Chromatographie, IR- und NMR-Spektroskopie. Die Klassifizierung der Pollenspezies erfolgte durch eine Clusteranalyse der Raman- Spektren. Diese Methode kann die Grundlage für eine automatische On-line-Detektion zur Erstellung von Pollenflugwarnungen für Allergiker bilden. Es gelang die Pollenarten zu unterscheiden, darüber hinaus spiegeln die resultierenden Dendrogramme den phylogenetischen Ursprung der Pollenspezies wider. Eine Ursache für falsche Zuordnungen der Pollen stellten hohe Gehalte an Carotinoiden dar. Durch das Bestrahlen der Proben mit 633 nm wurden die Carotinoide abgebaut und der Einfluss der Carotinoide auf die Clusteranalyse reduziert. Die Aufnahmen von resonanten Raman-Spektren der mittels HPTLC aufgetrennten Carotinoide belegten Unterschiede in der Zusammensetzung der Carotinoide in Abhängigkeit von der Pollenspezies. Die Konzentrationen der Carotinoide in den Pollen konnten mit Hilfe der UV-Vis Absorptions- Spektroskopie bestimmt werden. Des Weiteren konnte belegt werden, dass die in-situ Spektren ganzer Pollenkörner die typischen Carotinoidsignaturen einer Pollenspezies widergeben. Die in-situ Spektren sind von Vorteil, weil sie ohne Präparation aufgenommen wurden. Außerdem wurde der Einfluss der biologischen Matrix berücksichtigt. Weitere Bestandteile der Pollenkörner wurden isoliert und mit Hilfe der Raman-, IR- und NMR-Spek- troskopie untersucht. So konnten Veränderungen der chemischen Struktur durch die Aufreinigungsprozeduren nachgewiesen werden. Von besonderem Interesse ist die Analyse des Biomakromoleküls Sporopollen in, welches für die Langlebigkeit und chemischen Stabilität der Pollenkörner verantwortlich ist. Es wurden Aminosäuren, Phosphorlipide, Cellulose und Coumarinsäure in den Pollenkörnern gefunden. Die Verteilung dieser Substanzen in den Pollenkörnern konnte via 2D Raman-Experimente dargestellt werden. Die Signale der Coumarinsäure konnten ebenfalls in den Raman-Spektren keimender Pollenkörner entdeckt werden. Die Pollenschläuche bestehen überwiegend aus Cellulose. Neben diesen Gemeinsamkeiten wurden speziesspezifische Unterschiede der chemischen Zusammensetzung festgestellt. N2 - The chemical composition of pollen, the physiological containers that produce the male gametophytes of seed plants, has been a subject of research of plant physiologists, biochemists, and lately even material scientists for various reasons. The aim of this work was the analysis of whole pollen grains and pollen components by Raman Spectroscopy. These experiments were complemented by other techniques such as Enviromental Scanning Electron Microscopy (ESEM), High-Performance-Thin-Layer-Chromatography (HPTLC), Infrared Spectroscopy (IR) and Nuclear-Magnetic-Resonance Spectroscopy (NMR). As reported here, individual fresh pollen grains and their morphological constituents can be characterized and also classified in situ without prior preparation. Classification of pollen is based on their biochemical fingerprint revealed in their Raman spectrum. Raman spectroscopy is nondestructive and can be carried out with single pollen grains or fragments. It could be shown that the biochemical makeup of the pollen (as a part of the recognition/mating system) is altered during formation of a new biological species and that the species-specific chemical similarities and dissimilarities indeed reflect in the Raman spectral fingerprint. On the basis of the chemical information, unsupervised multivariate analysis consisting of hierarchical clustering revealed in most cases chemical similarities between species that were indicative of both phylogenetic relationship and matin behavior. Therefore experiments were conducted that gave the in situ Raman spectroscopic signatures ot the carotenoid molecules. As the data indicates, the in situ Raman spectra of the carotenoid molecules measured in single intact pollen grains provide in situ evidence of interspecies variations in pollen carotenoid content, structure, and/or assembly without composition varied greatly between species and that the different in situ spectral signatures reflect indeed differences in carotenoid composition. Moreover, comparison of the HPTLC data with resonant Raman spectra measured in situ illustrated that resonant Raman spectroscopy is very sensitive compared to approaches that need to rely on extraction procedures. In addition, the in situ difference spectra already include spectral features that may arise due to association of the carotenoid molecules with the biological matrix. A database of the species-specific in situ carotenoid spectra could in the future be used for automated spectrum correction and identification. This would fulfill an important prerequisite for fast analysis of pollen, and also of other plant tissues, e.g., in quality control of produce. To learn more about components present in pollen other than carotenoids Raman, IR and NMR spectra were collected from purified pollen compounds. Structural changes due to the applied isolation procedures could be detected. This fact has to be kept in mind for the structural elucidation of sporopollenin. This macromolecule is responsible for the longevity and stability of pollen, hence possible applications in materials science palynology and food technology arise. Based on the Raman spectra numerous compounds were detected within pollen such as amino acids, phospholipids, cellulose and coumaric acid. The distribution of these molecules within the pollen grains was illustrated by 2D Raman mapping experiments. Apart from microspectroscopic studies of quiescent, inactive pollen grains, also the dynamic process of pollen germination was investigated by Raman spectroscopy. The result permits the conclusion that differences between the spectra of germinated and non germinated pollen are caused by metabolic changes. This shows that Raman spectroscopy is not only a tool for fingerprinting and structure analysis but could also be useful for other disciplines. T3 - BAM Dissertationsreihe - 57 KW - Raman-Spektroskopie KW - Carotinoid KW - Clusteranalyse KW - Pollen KW - Sporopollenin PY - 2010 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:b43-1010 SN - 978-3-9813346-8-5 SN - 1613-4249 VL - 57 SP - 1 EP - 189 PB - Bundesanstalt für Materialforschung und -prüfung (BAM) CY - Berlin AN - OPUS4-101 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schulte, Franziska A1 - Kneipp, Janina A1 - Panne, Ulrich T1 - Vibrational spectroscopic characterization an identification of Pollen T2 - ECSBM 2007 CY - Paris, France DA - 2007-09-01 PY - 2007 AN - OPUS4-17983 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schulte, Franziska A1 - Panne, Ulrich A1 - Kneipp, Janina T1 - Characterisation of Pollen and their Constituents by Raman Spectroscopy T2 - ICORS CY - London, England DA - 2008-08-17 PY - 2008 AN - OPUS4-18083 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Schulte, Franziska A1 - Brzezinka, K.-W. A1 - Lutzenberger, K. A1 - Stege, H. A1 - Panne, Ulrich T1 - Raman spectroscopy of synthetic organic pigments used in 20th century works of art N2 - Raman microscopy allows a non-destructive characterisation of inorganic and organic painting materials such as pigments and organic dyestuffs. The objectives of this study are the more recent organic pigments typically present in paintings and other art works from the 20th century. More than 20 organic synthetic pigments from different chemical classes could be identified by Raman spectroscopy using different excitation wavelengths (457.9, 476.5, 487.9, 514.5, 632.8, and 1064 nm). To evaluate the performance for real paint samples, varying paint mixtures of the Hansa Yellow pigment PY 3 and the binding medium Mowilith, a polyvinyl acetate (PVAC) compound, were characterised; PY 3 was determined at a 1 wt% level in the binder. In addition, commercial tube paints containing the quinacridone violet PV 19 were studied. The pigment was clearly identified in all of these more complex oil and acrylic paints. Finally, alizarin (PR 83) and a green copper phthalocyanine pigment (PG 7) could unambiguously be identified by Raman microscopy in the painting Woman with mandolin in yellow and red of Max Beckmann dating 1950. The discovery of a red naphthol AS pigment by Raman spectroscopy in a sample from the Three field workers by Georg Baselitz (1964/1965) demonstrated that in some cases complementary chromatographic methods are needed for a comprehensive identification of the organic pigments. KW - Raman microscopy KW - FT-Raman spectroscopy KW - Synthetic organic pigments KW - Modern art PY - 2008 U6 - https://doi.org/10.1002/jrs.2021 SN - 0377-0486 SN - 1097-4555 VL - 39 IS - 10 SP - 1455 EP - 1463 PB - Wiley CY - Chichester AN - OPUS4-18401 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schulte, Franziska A1 - Panne, Ulrich A1 - Kneipp, Janina T1 - Raman-Spektroskopie als potentielles Werkzeug zur on-line Identifizierung von Pollen T2 - 4. Kolloquium Arbeitskreises Prozessanalytik der GDCh CY - Basel, Switzerland DA - 2008-11-17 PY - 2008 AN - OPUS4-18375 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schulte, Franziska A1 - Lutzenberger, K. A1 - Brezinka, K.W. A1 - Stege, H. A1 - Panne, Ulrich T1 - Raman Microscopy on Organic Synthetic Pigments- from Pure References to Painting Samples T2 - Scientific workshop on organic synthetic colourants CY - Munich, Germany DA - 2008-09-12 PY - 2008 AN - OPUS4-18080 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schulte, Franziska A1 - Brzezinka, K. W. A1 - Stege, H. A1 - Panne, Ulrich T1 - Analysis of synthetic organic pigments used in work of art and commercial paints by Raman spectroscopy T2 - Euroanalysis CY - Antwerp, Belgium DA - 2007-09-11 PY - 2007 AN - OPUS4-18081 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schulte, Franziska A1 - Panne, Ulrich A1 - Kneipp, Janina T1 - Classification and Indentification of Pollen by Raman Spectroscopy T2 - 12th International Palynological Congress (IPC-XII 2008) CY - Bonn, Germany DA - 2008-08-30 PY - 2008 AN - OPUS4-18082 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schulte, Franziska A1 - Lutzenberger, K. A1 - Brzezinka, K.-W. A1 - Stege, H. A1 - Panne, Ulrich T1 - Raman Microscopy on Organic Synthetic Pigments from Pure References to Painting Samples T2 - Scientific workshop on organic synthetic colourants CY - Munich, Germany DA - 2008-09-12 PY - 2008 AN - OPUS4-18491 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Schulte, Franziska A1 - Lingott, J. A1 - Panne, Ulrich A1 - Kneipp, Janina T1 - Chemical characterization and classification of Pollen N2 - We report on the in situ characterization of tree pollen molecular composition based on Raman spectroscopy. Different from purification-based analysis, the nondestructive approach allows (i) to analyze various classes of molecules simultaneously at microscopic resolution and (ii) to acquire fingerprint-like chemical information that was used for the classification of pollen from different species. Hierarchical cluster analysis of spectra from fresh pollen samples of 15 species partly related at the genus level and family level indicates separation of species based on the complete Raman spectral signature and yields classification in accord with biological systematics. The results have implications for the further elucidation of pollen biochemistry and also for the development of chemistry-based online pollen identification methods. PY - 2008 U6 - https://doi.org/10.1021/ac801791a SN - 0003-2700 SN - 1520-6882 SP - 1 EP - 12 PB - American Chemical Society CY - Washington, DC AN - OPUS4-18234 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schulte, Franziska A1 - Panne, Ulrich A1 - Kneipp, Janina T1 - Characterization and Identification of Pollen by Raman Spectroscopy T2 - ANAKON CY - Berlin, Germany DA - 2009-03-17 PY - 2009 AN - OPUS4-19208 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schulte, Franziska A1 - Panne, Ulrich A1 - Kneipp, Janina T1 - Klassifizierung von Pollen anhand ihrer Raman-Spektren: Auf dem Weg zur On-line-Pollenwarnung T2 - Doktorandentreffen CY - Attendorn, Germany DA - 2009-02-15 PY - 2009 AN - OPUS4-19203 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Schulte, Franziska A1 - Panne, Ulrich A1 - Kneipp, Janina T1 - Molecular changes during pollen germination can be monitored by Raman microspectroscopy N2 - The processes associated with pollen germination were studied in vitro for two tree species, Salix caprea and Fraxinus excelsior under different nutrient conditions. The results provide evidence of changes in chemical composition of the pollen grains during germination. From the comparison of spectra of the pollen grain body and the growing pollen tube, it can be concluded that there are major chemical differences between these two morphological units. Comparison of germinated and ungerminated pollen grains reveals alterations in the metabolism. Composition of the germinating pollen grain and its morphological units depends on the plant species, but also on the nutrient conditions. The results suggest species-specific utilization of metabolite storage, and potential alterations of the pollen outer coat. Furthermore, discharge of molecules into the nutrient medium may depend on the nutrient conditions in the germination experiments. This has implications for further experiments on dynamic processes in pollen and related plant materials. KW - Pollen germination KW - Raman microspectroscopy KW - Imaging KW - Coumaric acid KW - Pollen tube KW - Sporopollenin PY - 2010 U6 - https://doi.org/10.1002/jbio.201000031 SN - 1864-063X VL - 3 IS - 8-9 SP - 542 EP - 547 PB - Wiley-VCH Verl. CY - Weinheim AN - OPUS4-23211 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schulte, Franziska A1 - Kneipp, Janina A1 - Panne, Ulrich A1 - Kroh, L. W. A1 - Mäder, J. T1 - HPTLC-supported and in-situ resonance Raman spectroscopy of carotenoid signature in pollen T2 - ECSBM CY - Palermo, Italy DA - 2009-08-28 PY - 2009 AN - OPUS4-19542 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schulte, Franziska A1 - Mäder, J. A1 - Kroh, L. W. A1 - Panne, Ulrich A1 - Kneipp, Janina T1 - Analysis of Carotenoids Contained in Pollen by Raman Spectroscopy T2 - Tag der Chemie CY - Potsdam, Germany DA - 2009-06-04 PY - 2009 AN - OPUS4-19304 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schulte, Franziska A1 - Kneipp, Janina A1 - Panne, Ulrich A1 - Kroh, L. W. A1 - Mäder, J. T1 - HPTLC-supported and in-situ resonance Raman spectroscopy of carotenoid signature in pollen T2 - Workshop, Robert-Koch-Institut CY - Berlin, Germany DA - 2009-10-15 PY - 2009 AN - OPUS4-20127 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Merk, Virginia A1 - Schulte, Franziska A1 - Rooch, Heidemarie A1 - Feldmann, Ines A1 - Dörfel, Ilona A1 - Österle, Werner A1 - Panne, Ulrich A1 - Kneipp, Janina T1 - Surface-enhanced Raman scattering with silver nanostructures generated in situ in a sporopollenin biopolymer matrix N2 - Silver nanoparticles were generated based on citrate reduction in the ultrastructure of the sporopollenin biopolymer of Ambrosia artemisiifolia (ragweed) and Secale cereale (rye). The nanoparticles enable the acquisition of SERS spectra and thereby a vibrational characterization of the local molecular structure of sporopollenin. PY - 2011 U6 - https://doi.org/10.1039/c0cc05326k SN - 0022-4936 SN - 0009-241x SN - 1359-7345 SN - 1364-548x N1 - Geburtsname von Merk, Virginia: Joseph, V. - Birth name of Merk, Virginia: Joseph, V. VL - 47 IS - 11 SP - 3236 EP - 3238 PB - Royal Society of Chemistry CY - Cambridge AN - OPUS4-23482 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Schulte, Franziska A1 - Mäder, J. A1 - Kroh, L.W. A1 - Panne, Ulrich A1 - Kneipp, Janina T1 - Characterization of pollen carotenoids with in situ and high-performance thin-layer chromatography supported resonant Raman spectroscopy N2 - Raman signatures of the carotenoid component are studied in individual pollen grains from different species of trees. The information is obtained as differences in the strong pre-resonant Raman spectra measured before and after photodepletion of the carotenoid molecules. The results provide the first in situ evidence of interspecies differences in pollen carotenoid content, structure, and/or assembly between plant species without prior purification. The analysis of carotenoids in situ is confirmed by high-performance thin-layer chromatography (HPTLC)-supported resonance Raman data measured directly on the HPTLC plates after separation of carotenoids in pollen extracts. Utilization of the in situ, extraction-free procedure in carotenoid analysis will improve sensitivity and structural selectivity and provides insight into carotenoid structure and composition in single pollen grains. PY - 2009 U6 - https://doi.org/10.1021/ac901389p SN - 0003-2700 SN - 1520-6882 VL - 81 IS - 20 SP - 8426 EP - 8433 PB - American Chemical Society CY - Washington, DC AN - OPUS4-20260 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -