Electrochemical Detection; Double-pulse LIBS; Laser-induced breakdown spectroscopy; Fluorescence Microscopy; Elemental Distribution; Mass Spectrometry;

Laser-Induced Breakdown Spectroscopy (LIBS) in double pulse configuration (DP LIBS) was used for scanning elemental spatial distribution in annual terminal stems of spruce (Picea abies (L.) Karsten). Cross sections of stems cultivated in Cu2+ solution of different concentrations were prepared and analyzed by DP LIBS. Raster scanning with 150 µm spatial resolution was set and 2D (2-dimentional) maps of Cu and Ca distribution were created on the basis of the data obtained. Stem parts originating in the vicinity of the implementation of the cross sections were mineralized and subsequently Cu and Ca contents were analyzed by inductively coupled plasma mass spectrometry (ICP-MS). The results provide quantitative information about overall concentration of the elements in places, where LIBS measurements were performed. The fluorescence pictures were created to compare LIBS distribution maps and the fluorescence intensity (or the increase in autofluorescence) was used for the comparison of ICP-MS quantitative results. Results from these three methods can be utilized for quantitative measurements of copper ions transport in different plant compartments in dependence on the concentration of cultivation medium and/or the time of cultivation.

Laser-induced breakdown spectroscopy; LIBS; Mapping; Trace elements; Phytoremediation; Cancer; Plant; Bacteria; Animal tissues;

Laser-Induced Breakdown Spectroscopy (LIBS) is a sensitive optical technique capable of fast multi-elemental analysis of solid, gaseous and liquid samples. Since the late 1980s LIBS became visible in the analytical atomic spectroscopy scene; its applications having been developed continuously since then. In this paper, the use of LIBS for trace element determination in different matrices is reviewed. The main emphasis is on spatially resolved analysis of microbiological, plant and animal samples.

Laser-induced breakdown spectroscopy; LIBS; imaging techniques; nanoparticles; fluorescence; magnetic resonance imaging;

The requirements for early diagnostics as well as effective treatment of insidious diseases such as cancer constantly increase the pressure on development of efficient and reliable methods for targeted drug/gene delivery as well as imaging of the treatment success/failure. One of the most recent approaches covering both the drug delivery as well as the imaging aspects is benefitting from the unique properties of nanomaterials. Therefore a new field called nanomedicine is attracting continuously growing attention. Nanoparticles, including fluorescent semiconductor nanocrystals (quantum dots) and magnetic nanoparticles, have proven their excellent properties for in vivo imaging techniques in a number of modalities such as magnetic resonance and fluorescence imaging, respectively. In this article, we review the main properties and applications of nanoparticles in various in vitro imaging techniques, including microscopy and/or laser breakdown spectroscopy and in vivo methods such as magnetic resonance imaging and/or fluorescence-based imaging. Moreover the advantages of the drug delivery performed by nanocarriers such as iron oxides, gold, biodegradable polymers, dendrimers, lipid based carriers such as liposomes or micelles are also highlighted.

Laser Induced Breakdown Spectroscopy (LIBS); Picea abie; heavy metals;

This work is focused on application of LIBS technique for the study of plant samples. The elemental mapping on cross section throughout the annual terminal twigs of Picea abies was performed using double pulse Laser Induced Breakdown Spectroscopy (DP LIBS). 2D maps were created, where distribution of Cu and Ca in the plant tissue was observed. After mineralization of twig parts originated in the vicinity of the implementation of cross section, ICP-MS analysis was used for determination of total content of investigated elements.

laser induced breakdown spectroscopy; heavy meta;, spatial distribution; phytoremediation; soil; plant;

In numerous biological experiments, there is no information about the spatial distribution of elements, especially metals, which could further serve for understanding the biochemical mechanism of their distribution and transportation. Laser induced breakdown spectroscopy (LIBS) is a useful technique for determining the spatial distribution of metals in various types of samples. Given the potential of this technique to analyze solid, liquid and gaseous samples (including aerosols), which may or may not be conductive, LIBS is used for both qualitative and quantitative purposes in many areas. In some applications, the unique properties of this technique, as the ability of micro-analysis, the possibility of analyzing a sample at a distance and in situ and quasi-non-destructive nature of analysis, are used. In this study, the basic principle of technology and its application for analysis of soils and plant tissues are discussed.

phytoremediation; heavy metals; sunflower; lead ions; high performance liquid chromatography with electrochemical detection; spectrometry; laser induced breakdown spectroscopy;

In this study, the influence of lead (II) ions on sunflower growth and biochemistry was investigated from various points of view. Sunflower plants were treated with 0, 10, 50, 100 and/or 500 µM Pb-EDTA for eight days. We observed alterations in growth in all experimental groups compared with non-treated control plants. Further we determined total content of proteins by a Bradford protein assay. By the eighth day of the experiment, total protein contents in all treated plants were much lower compared to control. Particularly noticeable was the loss of approx. 8 µg/mL or 15 µg/mL in shoots or roots of plants treated with 100 mM Pb-EDTA. We also focused our attention on the activity of alanine transaminase (ALT), aspartate transaminase (AST) and urease. Activity of the enzymes increased with increasing length of the treatment and applied concentration of lead (II) ions. This increase corresponds well with a higher metabolic activity of treated plants. Contents of cysteine, reduced glutathione (GSH), oxidized glutathione (GSSG) and phytochelatin 2 (PC2) were determined by high performance liquid chromatography with electrochemical detection. Cysteine content declined in roots of plants with the increasing time of treatment of plants with Pb-EDTA and the concentration of toxic substance. Moreover, we observed ten times higher content of cysteine in roots in comparison with shoots. The observed reduction of cysteine content probably relates with its utilization for biosynthesis of GSH and phytochelatins, because the content of GSH and PC2 was similar in roots and shoots and increased with increased treatment time and concentration of Pb-EDTA. Moreover, we observed oxidative stress caused by Pb-EDTA in roots where the GSSG/GSH ratio was about 0.66. In shoots, the oxidative stress was less distinctive, with a GSSG/GSH ratio 0.14. We also estimated the rate of phytochelatin biosynthesis from the slope of linear equations plotted with data measured in the particular experimental group. The highest rate was detected in roots treated with 100 µM of Pb-EDTA. To determine heavy metal ions many analytical instruments can be used, however, most of them are only able to quantify total content of the metals. This problem can be overcome using laser induced breakdown spectroscopy, because it is able to provide a high spatial-distribution of metal ions in different types of materials, including plant tissues. Data obtained were used to assemble 3D maps of Pb and Mg distribution. Distribution of these elements is concentrated around main vascular bundle of leaf, which means around midrib.

Metallomics and metalloproteomics are emerging fields addressing the role, uptake, transport and storage of trace metals essential for life. There are several main approaches that are being developed in metallomics and metalloproteomics including both detection of levels and spatial distribution of heavy metals and determination of heavy-metal-contained proteins. The aim of this work is detection of copper and zinc in healthy and tumour tissues of miniature pigs by using of the laser induced breakdown spectroscopy (LIBS). Concentration of heavy metal transporting protein metallothionein (MT) was determined by Brdicka reaction. Tissue cryosections were obtained from the MeLiM strain of miniature pigs with hereditary melanoma, particularly from healthy skin, cutaneous nodular melanomas and metastases in the liver, spleen and lymph nodes. Using LIBS we measured maps of spatial distribution of the essential heavy metals in cryosections and found that the maps of healthy and tumour cryosection markedly differed. The highest content of MT was determined in the tumours localised on the back of animals and was nearly 500 ug of MT per gram of tissue.

Silver; Heavy metals; Plant biosensor; Sensors; Biochemical marker;

The aim of this work is to investigate sunflower plants response on stress induced by silver(I) ions. The sunflower plants were exposed to silver(I) ions (0, 0.1, 0.5, and 1 mM) for 96 h. Primarily we aimed our attention to observation of basic physiological parameters. We found that the treated plants embodied growth depression, coloured changes and lack root hairs. Using of autofluorescence of anatomical structures, such as lignified cell walls, it was possible to determine the changes of important shoot and root structures, mainly vascular bungles and development of secondary thickening. The differences in vascular bundles organisation, parenchymatic pith development in the root centre and the reduction of phloem part of vascular bundles were well observable. Moreover with increasing silver(I) ions concentration the vitality of rhizodermal cells declined; rhizodermal cells early necrosed and were replaced by the cells of exodermis. Further we employed laser induced breakdown spectroscopy for determination of spatial distribution of silver(I) ions in tissues of the treated plants. The Ag is accumulated mainly in near-root part of the sample. Moreover basic biochemical indicators of environmental stress were investigated. The total content of proteins expressively decreased with increasing silver(I) ions dose and the time of the treatment. As we compare the results obtained by protein analysis-the total protein contents in shoot as well as root parts-we can assume on the transport of the proteins from the roots to shoots. This phenomenon can be related with the cascade of processes connecting with photosynthesis. The second biochemical parameter, which we investigated, was urease activity. If we compared the activity in treated plants with control, we found out that presence of silver(I) ions markedly enhanced the activity of urease at all applied doses of this toxic metal. Finally we studied the effect of silver(I) ions on activity of urease in in vitro conditions.

Laser induced Breakdown Spectroscopy (LIBS) offers a new unique tool to detect heavy metal ions in biological samples without any sample preparation. The technique was utilized for determination of lead in leaves of sugar beet plants treated with lead chelate.