Generation of luminescent materials through living micoalgae

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In this follow-up project we want to further investigate biomineralization processes in the marine alga Emiliania huxleyi. We want to reveal the pathway of the lanthanide terbium from the surrounding medium into the intracellular coccolith vesicle (cv), where the coccolith is produced. The incorporation of Tb3+ into the coccoliths was proven in part 1 of the project, these results showed, that Tb3+ is incorporated into the growing calcite coccolith in the cv inside the cells. The advantage of investigating the pathway of Tb3+ through the cells is its specific fluorescence when excited with light. For the investigation of the transport and pathway of Tb3+ in the cell, we want to use two different approaches: (A), we want to identify the proteins which are involved in the transportation of Tb through the cell using 2D electrophoresis. Here we want to compare the proteomes of the calcifying E. huxleyi strain CCMP1516 with the non-calcifying strain CCMP1516NC and at different amounts of Tb3+ and/or Ca2+ in the medium. (B), we want to reveal in which organelles and/or compartments of the cells the Tb is accumulated during its way from the cell membrane to the cv by Confocal microscopy and Electron microscopy techniques. First, we will observe the living cells after adding Tb3+ to the medium using a confocal microscope. We want to track the distribution of the Tb3+ in the cells using a filter, thereby we can visualize exclusively the Tb3+ emission. Since the formation of the intracellular coccoliths occurs within 1h to 2h, we will evaluate, where inside the cells the Tb3+ is accumulated during this time period. When we can observe its accumulation at a specific site in the cells, the corresponding cells will be sampled and further treated for the investigation by TEM under cryo-conditions, i.e. first freezing the cells with high pressure freezer (HPF), cryo-substitute them and after cutting analyze them using Cryo TEM/EDX and a TEM equipped with CL.