Three dimensional SR-confocal micro XRF combined with absorption microtomography on Daphnia magna: Instrumental developments and recent advances in dataprocessing
Abstract
Daphnia magna are being used to study the mechanism of intoxication by metals (Zn, Cu, Ni) and to develop so called biotic ligand models (BLM's). It has been shown that Daphnia magna responds differently to Zn-exposure via the water and/or via the food. In order to distinguish critical tissue from whole body accumulation in these small organisms (3mm size), we explored the potential of combined X-ray techniques as a complementary approach to the above problem. Fast dynamic 2D micro XRF scanning and XRF micro CT experiments on Daphnia magna at Beamline L (HASYLAB, Hamburg, Germany) allowed the semi-quantitative investigation of the accumulation of metals within specific organs with microscopic resolution. Combining these results with micro CT data sets unravelled the tissue specifi 2D/3D distribution of metals within delicate organic sample on the 3-15 mu resolution revel in an essentially non-destructive manner. As demonstrated, recent developments at beamline L, involving a dedicated confocal micro-XRF set-up with a resolution of 5 mu and full 3D dynamic scanning algorithms allowed to explore microscopic 3D substructures within Daphnia magna (also called "top-down" methodology) aiding ecotoxicological research towards the tissue and even cellular level. Three dimensional isosurfaces of the elements of interest were obtained within a 300 mu Daphnia magna egg. Moreover, K-means clustering allowed us to isolate 3D clusters of interest of which the elemental content can be…
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