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Raman microspectra combine information on chemical composition of plant tissues with spatial information. The contributions from the building blocks of the cell walls in the Raman spectra of plant tissues can vary in the microscopic sub-structures of the tissue. Here, we discuss the analysis of 55 Raman maps of root, stem, and leaf tissues of Cucumis sativus, using different spectral contributions from cellulose and lignin in both univariate and multivariate imaging methods. Imaging based on hierarchical cluster analysis (HCA) and principal component analysis (PCA) indicates different substructures in the xylem cell walls of the different tissues. Using specific signals from the cell wall spectra, analysis of the whole set of different tissue sections based on the Raman images reveals differences in xylem tissue morphology. Due to the specifics of excitation of the Raman spectra in the visible wavelength range (532 nm), which is, e.g., in resonance with carotenoid species, effects of photobleaching and the possibility of exploiting depletion difference spectra for molecular characterization in Raman imaging of plants are discussed. The reported results provide both, specific information on the molecular composition of cucumber tissue Raman spectra, and general directions for future imaging studies in plant tissues.
Expansion dynamics of laser-induced plasma is studied for different focal positions of the ablation laser in the pressure range 10-2 - 105 Pa of the ambient air. The experimental results indicate that both the parameters significantly affect the plasma size, shape, intensity, reproducibility, and distance from the target surface. At pressures above 10 Pa, the plasma plume is confined by the ambient gas; the plumes are more compact and travel shorter distances from the target as compared to the analogous plume characteristics at pressures below 10 Pa. The pulse-to-pulse reproducibility of the integral emission intensity of the plasma is also different for different focal positions and pressures. It is found that the focal positions -1 cm and -2 cm below the target surface yield the most reproducible and intense emission signals as measured at the 600 ns delay time with the 100 ns gate. The information obtained can be of importance for pulsed laser deposition, laser welding, and analytical spectroscopy at reduced pressures. In general, a correct choice of the focal position and pressure of an ambient gas is very important for obtaining the strongest plasma emission, good reproducibility, and desired plasma plume shape.
Objective: The utilization of fluorescein-guided biopsies and resection has been recently discussed as a suitable strategy to improve and expedite operative techniques for the resection of central nervous system (CNS) tumors. However, little is known about the optical properties of sodium fluorescein (NaFl) in human tumor tissue and their potential impact on ex vivo analyses involving fluorescence-based methods.
Methods: Tumor tissue was obtained from a study cohort of an observational study on the utilization of fluorescein-guided biopsy and resection (n=5). The optical properties of fluorescein-stained tissue were compared to the optical features of the dye in vitro and in control samples consisting of tumor tissue of high-grade glioma patients (n=3) without intravenous (i.v.) application of NaFl. The dye-exposed tumor tissues were used for optical measurements to confirm the detectability of NaFl emission ex vivo. The tissue samples were fixed in 4%PFA, immersed in 30% sucrose, embedded in Tissue-Tek OCT compound, and cut to 10 mm cryosections. Spatially resolved emission spectra from tumor samples were recorded on representative slides with a Confocal Laser Scanning Microscope FV1000 (Olympus GmbH, Hamburg, Germany) upon excitation with lexc = 488 nm.
Results: Optical measurements of fluorescein in 0.9% sodium chloride (NaCl) under in vitro conditions showed an absorption maximum of lmax abs = 479 nm as detected with spectrophotometer Specord 200 and an emission peak at lmax em = 538 nm recorded with the emCCD detection system of a custom-made microscope-based single particle setup using a 500 nm long-pass filter. Further measurements revealed pH- and concentration-dependent emission spectra of NaFl. Under ex vivo conditions, confocal laser scanning microscopy of fluorescein tumor samples revealed a slight bathochromic shift and a broadening of the emission band.
Conclusion: Tumor uptake of NaFl leads to changes in the optical properties – a bathochromic shift and broadening of the emission band – possibly caused by the dye’s high pH sensitivity and concentration-dependent reabsorption acting as an innerfilter of the dye’s emission, particularly in the short wavelength region of the Emission spectrum where absorption and fluorescence overlap. Understanding the ex vivo optical properties of fluorescein is crucial for testing and validating its further applicability as an optical probe for intravital microscopy, immunofluorescence localization studies, and flow cytometry analysis.
In this article we demonstrate the use of atomic force microscopy (AFM) measurements for the study of macroscopic wear scars. By stitching AFM images acquired over the wear scar, the detailed structure of the scar can be characterized even when the scar is much wider than the typical maximum scan range of the AFM (50–100 µm). The results obtained by AFM are compared with those yielded by white light interferometry (WLI). The comparison validates the WLI measurements; at the same time, it shows decisive differences in the resolutions of these two methods. As a consequence, AFM measurements are necessary whenever a precise characterization of the structure of the scar is required. However, since stitching of AFM images is rather time-consuming, white light interferometry is recommended as a faster method whenever experiments are aimed at just a gross characterization of the scar and the measurement of mean quantities (e.g. the wear volume).
LIBS is a complementary method to XRF and can detect all elements without the need for vacuum conditions. Automated systems are already commercially available capable of scanning surfaces with a resolution of up to 0.1 mm within a few minutes. In addition to possible applications in R&D, LIBS is also used for practical applications in building materials laboratories and even on-site.
In view of ageing infrastructure facilities, a reliable assessment of the condition of concrete structures is of increasing interest. For concrete structures, the ingress of potential harmful ions is affecting the serviceability and eventually structural performance. Pitting corrosion induced by penetrating chlorides is the dominant deterioration mechanism. Condition assessment based on frequently performed chloride profiling can be useful to identify the extent and evolution of chloride ingress. This could prove to be more economical than extensive repairs, especially for important infrastructure facilities.
Currently the most common procedure for determining the chloride content is wet chemical analysis with standard resolution of 10 mm. The heterogeneity is not considered. LIBS is an economical alternative for determining the chloride content at depth intervals of 1 mm or less. It provides 2D distributions of multiple elements and can locate spots with higher concentrations. The results are directly correlated to the mass of binder and can also be performed on-site with a mobile LIBS-System.
The application of a LIBS-system is presented. Calibration is required for quantitative analysis. Concrete cores were drilled, sliced and analyzed to determine the 2D-distribution of harmful elements. By comparing the chloride ingress and the carbonation, the interaction of both processes can be visualized in a measurement that takes less than 10 minutes for a 50 mm x 100 mm drill core.
A leaflet on the use of LIBS for the chloride ingress assessment has been completed.
RATIONALE
The fast and univocal identification of different species in mixtures of pollen grains is still a challenge. Apart from microscopic evaluation and Raman spectroscopy, no other techniques are available.
METHODS
Matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry was applied to the analysis of extracts of single pollen grains and pollen mixtures. Pollen grains were fixed, treated and covered with matrix directly on the MALDI target.
RESULTS
Clearly resolved MALDI ion intensity images could be obtained enabling the identification of single pollen grains in a mixture.
CONCLUSIONS
Our results demonstrate the potential and the suitability of MALDI imaging mass spectrometry as an additional method for the identification of pollen mixtures.
A new approach that utilizes MALDI-TOF imaging mass spectrometry as a new detector for polymer chromatography is presented. For the first time, the individual retention behavior of single structural units of polyethylene oxide (PEO)/polypropylene oxide (PPO) copolymers and changes of the copolymer composition could be monitored. Composition specific calibration curves could be easily obtained by displaying the copolymer ion intensity data. This approach provides completely new insights in the chromatographic principle of copolymer separation and could be used to easily modify and adapt conditions for separation. In combination with electrospray deposition, homogeneous sample/matrix traces of surprisingly high spatial resolution could be obtained.
The localization of polymeric composition in samples prepared for matrix-assisted laser desorption/ionization (MALDI) analysis has been investigated by imaging mass spectrometry. Various matrices and solvents were used for sample spot preparation of a polybutyleneglycol (PBG 1000). It was shown that in visibly homogeneous spots, prepared using the dried droplet method, separation between matrix and polymer takes place. Moreover, using -cyano-4-hydroxycinnamic acid (CCA) as matrix and methanol as solvent molecular mass separation of the polymer homologues in the spots was detectable. In contrast to manually spotted samples, dry spray deposition results in homogeneous layers showing no separation effects.
We report on the use of a sprayable and thermogelating biomaterial (Poloxamer; a.k.a. Pluronic) in optical imaging of pH values, local oxygen and temperature. The material is highly biocompatible and easy to handle. We also show that the material is well permeable to oxygen (thus making it a good choice for use in oxygen sensors), and is stable in liquid solution and at elevated temperature. We demonstrate its applicability in optical sensors for oxygen, pH and temperature. This was accomplished by incorporating appropriate luminescent probes in various kinds of microparticles (which act as hosts for the probes and prevent dye leaching and aggregation), and then dispersing the microparticles in the thermogelating polymer. The resulting sensor gels were deposited on the surface of interest via spraying at temperatures of <20 °C. At these temperatures, the gels adhere well to the target, even on uneven surfaces such as skin, wounds, and bacterial cultures. If temperature is risen to above 25 °C, the gels form a thin and soft but solid sensing layer which, however, can be simply removed from surface of interest by cooling and wiping it off, or by washing with water. Sprayable thermogelating sensors present obvious advantages over other sensors by not causing damage to the surface of interest. In our perception, the sensing materials also have wide further applicability in sensors for other species including clinically relevant gases, enzyme substrates (such as glucose or lactate) and ions.
Basic principles and technical realization of Auger Electron Spectroscopy (AES) are presented. Different modes of application are briefly addressed. These are spectroscopy at nanoscaled surface layers including quantitative element analysis and chemical state analysis, in-depth element analysis, imaging and element mapping. A survey on standards related to application of AES in surface chemical analysis is given.