Contact X-ray microscopy of living cells by using LiF crystal as imaging detector.

In this paper, the use of lithium fluoride (LiF) as imaging radiation detector to analyse living cells by single-shot soft X-ray contact microscopy is presented. High resolved X-ray images on LiF of cyanobacterium Leptolyngbya VRUC135, two unicellular microalgae of the genus Chlamydomonas and mouse macrophage cells (line RAW 264.7) have been obtained utilizing X-ray radiation in the water window energy range from a laser plasma source. The used method is based on loading of the samples, the cell suspension, in a special holder where they are in close contact with a LiF crystal solid-state X-ray imaging detector. After exposure and sample removal, the images stored in LiF by the soft X-ray contact microscopy technique are read by an optical microscope in fluorescence mode. The clear image of the mucilaginous sheath the structure of the filamentous Leptolyngbya and the visible nucleolus in the macrophage cells image, are noteworthiness results. The peculiarities of the used X-ray radiation and of the LiF imaging detector allow obtaining images in absorption contrast revealing the internal structures of the investigated samples at high spatial resolution. Moreover, the wide dynamic range of the LiF imaging detector contributes to obtain high-quality images. In particular, we demonstrate that this peculiar characteristic of LiF detector allows enhancing the contrast and reveal details even when they were obscured by a nonuniform stray light.

Characterization of biofilm-forming cyanobacteria for biomass and lipid production.

AIMS: This work reports on one of the first attempts to use biofilm-forming cyanobacteria for biomass and lipid production. METHODS AND RESULTS: Three isolates of filamentous cyanobacteria were obtained from biofilms at different Italian sites and characterized by a polyphasic approach, involving microscopic observations, ecology and genetic diversity (studying the 16S rRNA gene). The isolates were grown in batch systems and in a semi-continuous flow incubator, specifically designed for biofilms development. Culture system affected biomass and lipid production, but did not influence the fatty acid profile. The composition of fatty acids was mainly palmitic acid (>50%) and less amounts of other saturated and monounsaturated fatty acids. Only two isolates contained two polyunsaturated fatty acids. CONCLUSIONS: Data obtained from the flow-lane incubator system would support a more economical and sustainable use of the benthic micro-organisms for biomass production. The produced lipids contained fatty acids suitable for a high-quality biodiesel production, showing high proportions of saturated and monounsaturated fatty acids. SIGNIFICANCE AND IMPACT OF THE STUDY: Data seem promising when taking into account the savings in cost and time derived from easy procedures for biomass harvesting, especially when being able to obtain the co-production of other valuable by-products.

1H-NMR analysis of water mobility in cultured phototrophic biofilms.

The present work reports on the first attempt to study water mobility in phototrophic biofilms, applying the (1)H-NMR relaxometry technique to closely monitored microbial communities grown in a microcosm under controlled ambient conditions. Longitudinal water proton relaxation times exhibited a bi-exponential behavior in all biofilm samples, indicating two types of water molecules with diverging dynamic properties, confined to different compartments of the biofilm. The fast-relaxing component can be attributed to water molecules tightly bound to the intracellular matrix, while the slow-relaxing component could reflect the behavior of water embedded in the biopolymer matrix, confined into matrix pores and channels. The results are discussed with respect to a possible key role of exopolysaccharides and uronic acids in water binding in phototrophic biofilms.

Capsular polysaccharides of cultured phototrophic biofilms.

Phototrophic biofilm samples from an Italian wastewater treatment plant were studied in microcosm experiments under varying irradiances, temperatures and flow regimes to assess the effects of environmental variables and phototrophic biomass on capsular exopolysaccharides (CPS). The results, obtained from circular dichroism spectroscopy and High Performance Liquid Chromatography, suggest that CPS have a stable spatial conformation and a complex monosaccharide composition. The total amount present was positively correlated with the biomass of cyanobacteria and diatoms, and negatively with the biovolume of green algae. The proportion of uronic acids showed the same correlation with these taxon groups, indicating a potential role of cyanobacteria and diatoms in the removal of residual nutrients and noxious cations in wastewater treatment. While overall biofilm growth was limited by low irradiance, high temperature (30 degrees C) and low flow velocity (25 l h(-1)) yielded the highest phototrophic biomass, the largest amount of CPS produced, and the highest proportion of carboxylic acids present.

A complex photoreceptive structure in the cyanobacterium Leptolyngbya sp.

Among the terrestrial epilithic cyanobacteria isolated from Roman hypogea at extremely low light intensity, a non-heterocystous strain, belonging to the genus Leptolyngbya, showed a marked photobehavior. These red cyanobacteria possess an orange spot at the tip of the apical cell. Micro-spectrophotometric analysis of this tip showed an absorption spectrum with two bands, centered at 456 and 504 nm, respectively. Experiments on photo-orientation impairment of these cells, and micro-spectrophotometric analysis of the tip of impaired trichomes showed that a rhodopsin-like protein might be present in this structure. All these data could support the hypothesis of the presence of a complex photoreceptive system in this prokaryote.

Structural Interactions among Epilithic Cyanobacteria and Heterotrophic Microorganisms in Roman Hypogea.

Abstract Phototrophic microbial communities present in the Roman Catacombs were characterized and different species of terrestrial epilithic cyanobacteria were found to occur as dominant organisms. Eucapsis, Leptolyngbya, Scytonema, and Fischerella were the most frequently encountered cyanobacterial taxa, while a few species of green algae and the diatom Diadesmis gallica occurred in minor amounts. Streptomyces strains, a few genera of eubacteria, and to a lesser extent fungi were always present in the same microhabitats and contributed to the deterioration of stone surfaces. The combined use of light and electron microscopy evidenced the structural relationships among rod-shaped or filamentous bacteria and cyanobacterial cells, as well as the presence of polysaccharide capsules and sheaths, and of mineral precipitates on S. julianum filaments. The significance of the intimate association among the microorganisms was discussed in relation to the damage caused by the growth of biological patinas on stone surfaces.http://link.springer-ny.com/link/service/journals/00248/bibs/38n3p244.html

Long-Duration Soft X-Ray Pulses by XeCl Laser Driven Plasmas and Applications.

We report the characterization of a soft x-ray plasma source generated by a long-pulse XeCl excimer laser system. The output energy is 4 J at a wavelength of 308 nm in a 100-ns pulse. The intensity of radiation on target is estimated to be 4 × 1012 W cm-2. X-ray emission spectra of the plasma have been recorded using a double focusing spatial resolution spectrometer with a spherical mica crystal. From these measurements, the plasma temperature and electron density have been estimated. Various applications of such a plasma source have been investigated. First images of whole intact living cells from our system, imaged using the technique of soft x-ray contact microscopy, utilizing x rays in the "water window" region (280-530 eV), are shown. The suitability of the source for other applications, for example, x-ray lithography and radiation damage studies, to living cells are discussed. Possible improvements to the x-ray source for the various applications are proposed.