Description: Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS) with a bismuth cluster primary ion source was used for analysing microbial lipid biomarkers in 10-µm-thick microscopic cryosections of methanotrophic microbial mats from the Black Sea. Without further sample preparation, archaeal isopranyl glycerol di- and tetraether core lipids, together with their intact diglycoside (gentiobiosyl-) derivatives, were simultaneously identified by exact mass determination. Utilizing the imaging capability of ToF-SIMS, the spatial distributions of these biomarkers were mapped at a lateral resolution of 〈 5 µm in 500 × 500 µm2 areas on the mat sections. Using inline image cluster projectiles in the burst alignment mode, it was possible to reach a lateral resolution of 1 µm on an area of 233 × 233 µm, thus approaching the typical size of microbial cells. The mappings showed different ‘provenances’ within the sections that are distinguished by individual lipid fingerprints, namely (A) the diethers archaeol and hydroxyarchaeol co-occurring with glycerol dialkyl glycerol tetraethers (GDGT), (B) hydroxyarchaeol and dihydroxyarchaeol, and (C) GDGT and gentiobiosyl-GDGT. Because ToF-SIMS is a virtually nondestructive technique affecting only the outermost layers of the sample surface (typically 10–100 nm), it was possible to further examine the studied areas using conventional microscopy, and associate the individual lipid patterns with specific morphological traits. This showed that provenance (B) was frequently associated with irregular, methane-derived CaCO3 crystallites, whereas provenance (C) revealed a population of fluorescent, filamentous microorganisms showing the morphology of known methanotrophic ANME-1 archaea. The direct coupling of imaging mass spectrometry with microscopic techniques reveals interesting perspectives for the in-situ study of lipids in geobiology, microbial ecology, and organic geochemistry. After further developing protocols for handling different kinds of environmental samples, ToF-SIMS could be used as a tool to attack many challenging problems in these fields, such as the attribution of biological source(s) to particular biomarkers in question, or the high-resolution tracking of biogeochemical processes in modern and ancient natural environments.

Description: Siliceous and calcareous sponges commonly are treated with acid to remove the spicules prior to embedding and cutting for histological investigations. Histology of spiculated sponge tissue represents a challenging problem in sponge histotechnology. Furthermore, fluorescence in situ hybridization (FISH), a key method for studying sponge-associated microbes, is not possible after acid treatment. For a broad range of siliceous sponge species, we developed and evaluated methods for embedding in paraffin, methylmethacrylate resins, LR White resin and cryomatrix. Different methods for cutting tissue blocks as well as mounting and staining sections also were tested. Our aim was to enable histological investigations and FISH without prior removal of the spicules. To obtain an overview of tissue and skeleton arrangement, we recommend embedding tissue blocks with LR White resin combined with en bloc staining techniques for large specimens with thick and numerous spicules, but paraffin embedding and subsequent staining for whole small specimens. For FISH on siliceous sponges, we recommend Histocryl embedding if the spicule content is high, but paraffin embedding if it is low. Classical histological techniques are used for detailed tissue examinations.

Description: Two different cyanobacterial biofilms from German karstwater creeks were investigated with respect to their photosynthetic effect on Ca2+ removal and potential CaCO3 precipitation in artificial creek waters of different CO2 partial pressures at a given, constant calcite supersaturation. CO2 partial pressures were adjusted to 350 ppmV, 2200 ppmV and 8700 ppmV respectively, covering the range of Phanerozoic atmospheric CO2 partial pressures inferred from palaeosoils, stomatal indices and model calculations. Microsensor measurements of calcium, pH and oxygen revealed differences in the potential to precipitate CaCO3 between the two model organisms Tychonema-relative strain SAG 2388 and Synechococcus sp. strain SAG 2387. Whereas a strong removal of Ca2+ from the solution was measured at Tychonema-relative biofilm, the Synechococcus sp. biofilm exercised a much lower Ca2+ removal during photosynthesis. Photosynthesis was enhanced in both organisms with increasing CO2 and HCO3−, as indicated by enhanced O2 production, but only for the motile filamentous taxon Tychonema-relative a concomitantly increasing calcium removal was measured. However, model calculations indicate that this short-term Ca2+ binding in the Tychonema-relative is due to complexation to exopolymers or oscillin, with no immediate CaCO3 precipitation. In contrast, Ca2+ and pH measurements at Synechococcus sp. biofilm could be consistent with immediate CaCO3 precipitation at the cells. In both biofilms, pH gradients increase with increasing pCO2 from 350 to 2200 ppmV due to enhanced photosynthesis, but decrease at a pCO2 of 8700 ppmV despite of further enhanced photosynthesis. This observation, regardless whether CO2 or HCO3− is used by the cyanobacteria, is in accordance with hydrochemical modeling demonstrating an increased DIC buffering at high pCO2 conditions. These results indicate that the potential of cyanobacteria to form spatially defined calcification pattern via pH gradients at their cell envelopes ('calcified cyanobacteria') increases at elevated pCO2, while at high pCO2 conditions Ca2+ binding and lowered pH microgradients lead to spatially diffuse calcification without defined cell envelope precipitates.

Description: Scientific drilling conducted at the inner slope of the Miocene central Ries impact crater recovered a partial section of crater lake sediments. Four sequences were recovered, composed of suevite-derived sandstones, thin lignite seams, bituminous shales, and marlstones to claystones. These flooding-evaporation sequences reflect the impact of short-term climatic fluctuations on a hydrologically closed basin. The superimposed trend from sequences rich in bituminous shales in the lower parts of the section to sequences dominated by organic-poor claystones and intercalated lignites in the upper parts of the section resembles that of the 300-m-thick central crater basin succession, which has previously been considered to reflect a climate-controlled development from an alkaline saline lake to a freshwater lake with temporary coal swamps. In the sediment core of Enkingen, however, the change from bituminous shales to organic-poor claystones with intercalated lignites is associated with a general increase in salinity, as indicated by (1) palynomorphs, (2) increase in 13 C of the lipid biomarker archaeol (bis-O-phytanylglycerol), and (3) the occurrence of 13 C-enriched C 20 /C 25 -archaeol (O-phytanyl-O-sesterterpanylglycerol) specific to halophilic Archaea. In addition, the unidirectional trend in 87 Sr/ 86 Sr of carbonates, declining from ratios of Variscan basement rocks toward marine ratios, indicates a change from (1) weathering of crystalline rocks and suevite to (2) ejected Jurassic sediments (Bunte Breccia) in the catchment area as the major source of ion influx to the lake. From that trend, a change in lake water composition and a general increase in ion concentrations are inferred. These new results can be applied to a reassessment of major parts of the lacustrine succession of the Ries crater. We use these data to propose a new hypothetical model for the chemical and ecological evolution of the Ries crater lake: (1) After the establishment of a stratified brackish eutrophic soda lake due to silicate weathering and evaporation, the increasing influx of waters from the Bunte Breccia carbonate and authigenic silicate precipitation led to a mesotrophic halite lake with marine-like ion ratios and concentrations. (2) Further increase in ions, among them Mg 2+ and Sr 2+ , resulted in hypersaline conditions with gypsum precipitation, low primary production, and phreatic Sr-rich dolomitization in marginal carbonates. (3) The final, sudden change to oligotrophic freshwater conditions is explained by the formation of an outlet late in the lake history. We conclude that the chemical and ecological evolution of the Ries lake therefore appears to have been mainly controlled by the weathering history of the catchment area, with climate fluctuations causing superimposed cycles. Similarly, changes in terrestrial palynomorph associations may at least partly reflect a change in soil types in the catchment area, from fertile, moist soils on suevite to dry karst soils and soils on Bunte Breccia. These interpretations imply that the initial suevite blanket of the Ries crater was much more continuous and widespread than previously assumed.