Christian Seim

A compact Laboratory Transmission X-ray Microscope for the water window

In the water window (2.2 – 4.4 nm) the attenuation of radiation in water is significantly smaller than in organic material. Therefore, intact biological specimen (e.g. cells) can be investigated in their natural environment. In order to make this technique accessible to users in a laboratory environment a Full-Field Laboratory Transmission X-ray Microscope (L-TXM) has been developed. The L-TXM is operated with a nitrogen laser plasma source employing an InnoSlab high power laser system for plasma generation. For microscopy the Lyα emission of highly ionized nitrogen at 2.48 nm is used. A laser plasma brightness of 5 × 1011 photons/(s × sr × μm2 in line at 2.48 nm) at a laser power of 70 W is demonstrated. In combination with a state-of-the-art Cr/V multilayer condenser mirror the sample is illuminated with 106 photons/(μm2 × s). Using objective zone plates 35 – 40 nm lines can be resolved with exposure times < 60 s. The exposure time can be further reduced to 20 s by the use of new multilayer condenser optics and operating the laser at its full power of 130 W. These exposure times enable cryo tomography in a laboratory environment.

Synchrotron-based ν-XRF mapping and μ-FTIR microscopy enable to look into the fate and effects of tattoo pigments in human skin

The increasing prevalence of tattoos provoked safety concerns with respect to particle distribution and effects inside the human body. We used skin and lymphatic tissues from human corpses to address local biokinetics by means of synchrotron X-ray fluorescence (XRF) techniques at both the micro (μ) and nano (ν) scale. Additional advanced mass spectrometry-based methodology enabled to demonstrate simultaneous transport of organic pigments, heavy metals and titanium dioxide from skin to regional lymph nodes. Among these compounds, organic pigments displayed the broadest size range with smallest species preferentially reaching the lymph nodes. Using synchrotron μ-FTIR analysis we were also able to detect ultrastructural changes of the tissue adjacent to tattoo particles through altered amide I α-helix to β-sheet protein ratios and elevated lipid contents. Altogether we report strong evidence for both migration and long-term deposition of toxic elements and tattoo pigments as well as for conformational alterations of biomolecules that likely contribute to cutaneous inflammation and other adversities upon tattooing.

Reconstruction of confocal micro-X-ray fluorescence spectroscopy depth scans obtained with a laboratory setup.

Depth profiling with confocal micro-X-ray fluorescence spectroscopy (confocal micro-XRF) is a nondestructive analytical method for obtaining elemental depth profiles in the micrometer region. Up until now, the quantitative reconstruction of thicknesses and elemental concentration of stratified samples has been only possible with monochromatic, thus, synchrotron radiation. In this work, we present a new calibration and reconstruction procedure, which renders quantification in the laboratory feasible. The proposed model uses the approximation of an effective spot size of the optic in the excitation channel and relies on the calibration of the transmission of this lens beforehand. Calibration issues are discussed and validation measurements on thick multielement reference material and a stratified system are presented.

Old traces, read anew – ‘The Reading Hermit’ painting in the light of X-ray fluorescence

There exist several very similar looking versions of the painting ‘The Reading Hermit’, all allegedly painted by Rembrandt Harmenszoon van Rijn (approx. in ∼1630 A.D., Leiden). The classification of Rembrandts paintings, which were produced by Rembrandt himself, in his academy by his students and the ones being mere copies is a crucial and difficult task. We gathered background evidence and performed elemental analyses by non-destructive micro-X-ray fluorescence (micro-XRF) in order to elucidate the paintings provenance. Elemental distributions of Ca, Mn, Fe and Cu show that the painting was presumably changed during the painting process, which indicates, together with neutron autoradiography (NAR) investigations, that this version of ‘The Reading Hermit’ is not a copy.

Are metals involved in tattoo-related hypersensitivity reactions? A case report

Allergic reactions to tattoos are not uncommon. However, identification of the culprit allergen(s) remains challenging.

Laboratory full-field transmission x-ray microscopy

X-ray microscopy in the water window has become a valuable imaging tool for a wide field of applications with a resolution in the nanometer regime. The emergence and the development of laboratory based transmission X-ray microscopes (LTXM) can be of great benefit to users, since LTXM provides access to a method previously limited to synchrotron facilities only. In recent years, measuring times in the laboratory have been reduced to the point, where tomography of aqueous cryofixated samples has become feasible. We report on a laboratory full-field transmission X-ray microscope based on a laser induced plasma source located at the Berlin Laboratory for innovative X-ray Technologies. A short introduction on full-field X-ray microscopy in the water window is given. We demonstrate that, with a thin disk laser-system (TDL), which provides an average power of ~15 W a spatial resolution of Δx = 41 nm ± 3 nm (half-pitch) is feasible. An image of a diatom recorded at 15 W average laser power with a magnification of 1125x captured in 5 min is presented.

3D nanoscale imaging of biological samples with laboratory-based soft X-ray sources

In microscopy, where the theoretical resolution limit depends on the wavelength of the probing light, radiation in the soft X-ray regime can be used to analyze samples that cannot be resolved with visible light microscopes. In the case of soft X-ray microscopy in the water-window, the energy range of the radiation lies between the absorption edges of carbon (at 284 eV, 4.36 nm) and oxygen (543 eV, 2.34 nm). As a result, carbon-based structures, such as biological samples, posses a strong absorption, whereas e.g. water is more transparent to this radiation. Microscopy in the water-window, therefore, allows the structural investigation of aqueous samples with resolutions of a few tens of nanometers and a penetration depth of up to 10μm. The development of highly brilliant laser-produced plasma-sources has enabled the transfer of Xray microscopy, that was formerly bound to synchrotron sources, to the laboratory, which opens the access of this method to a broader scientific community. The Laboratory Transmission X-ray Microscope at the Berlin Laboratory for innovative X-ray technologies (BLiX) runs with a laser produced nitrogen plasma that emits radiation in the soft X-ray regime. The mentioned high penetration depth can be exploited to analyze biological samples in their natural state and with several projection angles. The obtained tomogram is the key to a more precise and global analysis of samples originating from various fields of life science.

Preparation of clay mineral samples for high resolution x-ray imaging

In the development of optimum ceramic materials for plastic forming, it is of fundamental importance to gain insight into the compositions of the clay minerals. Whereas spectroscopic methods are adequate for determining the elemental composition of a given sample, a knowledge of the spatial composition, together with the shape and size of the particles leads to further, valuable insight. This requires an imaging technique such as high resolution X-ray microscopy. In addition, fluorescence spectroscopy provides a viable element mapping technique. Since the fine particle fraction of the materials has a major effect on physical properties like plasticity, the analysis is focused mainly on the smallest particles. To separate these from the bigger agglomerates, the raw material has to pass through several procedures like centrifugation and filtering. After that, one has to deposit a layer of appropriate thickness on to a suitable substrate. These preparative techniques are described here, starting from the clay mineral raw materials and proceeding through to samples that are ready to analyze. First results using high resolution x-ray imaging are shown.