Characterization of Pollen Carotenoids with in situ and High-Performance Thin-Layer Chromatography Supported Resonant Raman Spectroscopy

Schulte, Franziska; Mäder, Jens; Kroh, Lothar W.; Panne, Ulrich; Kneipp, Janina

doi:10.1021/ac901389p

Cited by 82 publications

(61 citation statements)

References 37 publications

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“…Pollen grains of various species can vary quite a lot in grain shape and size, number, type and position of apertures, and structure and sculpture of the grain wall. In the last decade, vibrational spectroscopy of pollen has been performed in macroscopic mode on bulk samples, resulting in the clear identification of main chemical constituents (Zimmermann and Kohler 2014;Zimmermann 2010;Parodi et al 2013;Pappas et al 2003;Gottardini et al 2007;Schulte et al 2010;Schulte et al 2009;Schulte et al 2008;Ivleva et al 2005).…”

Section: Introductionmentioning

confidence: 99%

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Vibrational microspectroscopy enables chemical characterization of single pollen grains as well as comparative analysis of plant species based on pollen ultrastructure

Zimmermann

Bağcıoğlu

Sandt

et al. 2015

Planta

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Main conclusion: Chemical imaging of pollen by vibrational microspectroscopy enables characterization of pollen ultrastructure, in particular phenylpropanoid components in grain wall for comparative study of extant and extinct plant species.Keywords: FTIR microspectroscopy, Raman microspectroscopy, Pinales, imaging, cell wall. 2 SUMMARYA detailed characterization of conifer (Pinales) pollen by vibrational microspectroscopy is presented. The main problems that arise during vibrational measurements were scatter and saturation issues in Fourier transform infrared (FTIR), and fluorescence and penetration depth issues in Raman. Single pollen grains larger than approx. 15 µm can be measured by FTIR microspectroscopy using conventional light sources, while smaller grains may be measured by employing synchrotron light sources. Pollen grains that were larger than 50 µm were too thick for FTIR imaging since the grain constituents absorbed almost all infrared light. Chemical images of pollen were obtained on sectioned samples, unveiling the distribution and concentration of proteins, carbohydrates, sporopollenins and lipids within pollen substructures. The comparative analysis of pollen species revealed that, compared with other Pinales pollens, Cedrus atlantica has a higher relative amount of lipid nutrients, as well as different chemical composition of grain wall sporopollenin. The pre-processing and data analysis, namely extended multiplicative signal correction and principal component analysis, offer simple estimate of imaging spectral data and indirect estimation of physical properties of pollen. The vibrational microspectroscopy study demonstrates that detailed chemical characterization of pollen can be obtained by measurement of an individual grain and pollen ultrastructure. Measurement of phenylpropanoid components in pollen grain wall could be used, not only for the reconstruction of past environments, but for assessment of diversity of plant species as well. Therefore, analysis of extant and extinct pollen species by vibrational spectroscopies is suggested as a valuable tool in biology, ecology and palaeosciences.3

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Section: Introductionmentioning

confidence: 99%

“…The problem of strong fluorescence in the Raman spectra of pollen can be circumvented by photo-bleaching, i.e. photodepletion of the carotenoid molecules (Ivleva et al 2005;Schulte et al 2008;Schulte et al 2009). However, the process not only destructs the sample partially, it also significantly increases the spectral acquisition time.…”

Section: Introductionmentioning

confidence: 99%

Vibrational microspectroscopy enables chemical characterization of single pollen grains as well as comparative analysis of plant species based on pollen ultrastructure

Zimmermann

Bağcıoğlu

Sandt

et al. 2015

Planta

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“…As ice nucleation in clouds influences precipitation initiation and radiative forcing (Pruppacher and Klett, 1997;Lohmann et al, 2002;Storelvmo et al, 2011), it plays an important role for both climate and weather. Ice formation in clouds occurs either through homogeneous or heterogeneous ice nucleation.…”

Section: Introductionmentioning

confidence: 99%

“…The ice activity of the pollen washing waters could be attributed to extractable compounds with masses between 100 and 300 kDa, which was determined via size-selective filtration. It is known that the pollen surface is covered with huge amounts of compounds (e.g., Breiteneder et al, 1989;Clarke et al, 1979;Schulte et al, 2009), with many of them being easily extractable in water. Another source for pollen material is the interior of the pollen that can become accessible by the bursting of grains in water (Schäppi et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

Immersion freezing of birch pollen washing water

et al. 2013

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Abstract. Birch pollen grains are known to be ice nucleating active biological particles. The ice nucleating activity has previously been tracked down to biological macromolecules that can be easily extracted from the pollen grains in water. In the present study, we investigated the immersion freezing behavior of these ice nucleating active (INA) macromolecules. Therefore we measured the frozen fractions of particles generated from birch pollen washing water as a function of temperature at the Leipzig Aerosol Cloud Interaction Simulator (LACIS). Two different birch pollen samples were considered, with one originating from Sweden and one from the Czech Republic. For the Czech and Swedish birch pollen samples, freezing was observed to start at −19 and −17 • C, respectively. The fraction of frozen droplets increased for both samples down to −24 • C. Further cooling did not increase the frozen fractions any more. Instead, a plateau formed at frozen fractions below 1. This fact could be used to determine the amount of INA macromolecules in the droplets examined here, which in turn allowed for the determination of nucleation rates for single INA macromolecules. The main differences between the Swedish birch pollen and the Czech birch pollen were obvious in the temperature range between −17 and −24 • C. In this range, a second plateau region could be seen for Swedish birch pollen. As we assume INA macromolecules to be the reason for the ice nucleation, we concluded that birch pollen is able to produce at least two different types of INA macromolecules. We were able to derive parameterizations for the heterogeneous nucleation rates for both INA macromolecule types, using two different methods: a simple exponential fit and the Soccer ball model. With these parameterization methods we were able to describe the ice nucleation behavior of single INA macromolecules from both the Czech and the Swedish birch pollen.

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“…Therefore, compared with chemical characterization, it is more challenging to achieve pollen/ spore identification using specific Raman bands. For some studies of Raman spectra of pollens and spores the particles are laid on a substrate and spectra are recorded using a Raman spectrometer (e.g., [25,26]). In those studies, a Raman excitation laser beam was scanned across each micro-size (15-60 mm) pollen particle with a spatial resolution of 1 mm.…”

Section: Introductionmentioning

confidence: 99%

Photophoretic trapping-Raman spectroscopy for single pollens and fungal spores trapped in air

Wang

Pan

Hill

et al. 2015

Journal of Quantitative Spectroscopy and Radiative Transfer

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a b s t r a c tPhotophoretic trapping-Raman spectroscopy (PTRS) is a new technique for measuring Raman spectra of particles that are held in air using photophoretic forces. It was initially demonstrated with Raman spectra of strongly-absorbing carbon nanoparticles (Pan et al.[44] (Opt Express 2012)). In the present paper we report the first demonstration of the use of PTRS to measure Raman spectra of absorbing and weakly-absorbing bioaerosol particles (pollens and spores). Raman spectra of three pollens and one smut spore in a size range of 6.2-41.8 mm illuminated at 488 nm are shown. Quality spectra were obtained in the Raman shift range of 1600-3400 cm À 1 in this exploratory study. Distinguishable Raman scattering signals with one or a few clear Raman peaks for all four aerosol particles were observed within the wavenumber region 2940-3030 cm À 1 . Peaks in this region are consistent with previous reports of Raman peaks in the 1600-3400 cm À 1 range for pollens and spores excited at 514 nm measured by a conventional Raman spectrometer. Noise in the spectra, the fluorescence background, and the weak Raman signals in most of the 1600-3400 cm À 1 region make some of the spectral features barely discernable or not discernable for these bioaerosols except the strong signal within 2940-3030 cm À 1 . Up to five bands are identified in the three pollens and only two bands appear in the fungal spore, but this may be because the fungal spore is so much smaller than any of the pollens. The fungal spore signal relative to the air-nitrogen Raman band is approximately 10 times smaller than that ratio for the pollens. The five bands are tentatively assigned to the CH 2 symmetric stretch at 2948 cm À 1 , CH 2 Fermi resonance stretch at 2970 cm À 1 , CH 3 symmetric stretch at 2990 cm À 1 , CH 3 out-of-plane end asymmetric stretch at 3010 cm À 1 , and unsaturated ¼ CH stretch at 3028 cm À 1 . The two dominant bands of the up-to-five Raman bands in the 2940-3030 cm À 1 region have a consistent band spacing of 25 cm À 1 in all four aerosols. Finally we discuss improvements to the PTRS that should provide a system which can trap a higher fraction of particle types and obtain Raman spectra over a larger range (e.g., 200-3600 cm À 1 ) than those achieved here.Published by Elsevier Ltd.

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Characterization of Pollen Carotenoids with in situ and High-Performance Thin-Layer Chromatography Supported Resonant Raman Spectroscopy

Cited by 82 publications

References 37 publications

Vibrational microspectroscopy enables chemical characterization of single pollen grains as well as comparative analysis of plant species based on pollen ultrastructure

Vibrational microspectroscopy enables chemical characterization of single pollen grains as well as comparative analysis of plant species based on pollen ultrastructure

Immersion freezing of birch pollen washing water

Photophoretic trapping-Raman spectroscopy for single pollens and fungal spores trapped in air

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