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Schlierf Group - Protein-based biomineralization

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Super-Resolution in Diatoms

Super-resolution microscopy techniques based on single-molecule localization microscopy (SMLM) have become increasingly popular over the last few years to image in vivo structures with a spatial resolution around 20nm. In particular co-localization of enzymes and the resolution of structural organization of cytoskeletal elements have become possible and yielded new insights in numerous molecular mechanisms.  Protein localization in diatoms mostly relies on high-resolution electron-microscopy (EM) techniques or diffraction-limited fluorescence microscopy. SMLM offers now the possibility to shift fluorescence microscopy closer to the high-resolution EM regime while retaining in vivo like conditions. Here, we present a screening of photo-activatable and photo-switchable fluorescent proteins for super-resolution microscopy in diatoms. The fluorescent proteins covered a broad range of emission wavelengths and were also fused to a silica-embedded silaffin protein in Thalassiosira pseudonana. We found that three out of six proteins could successfully be activated or switched, while being embedded in the silica cell wall. The localization accuracy for the lateral resolution was in the range of 23nm ± 8nm allowing now direct visual access to the intricate protein-based nanopatterning of the diatom cell wall.

Team members on the project: Adeeba, Philip

Read more: Gröger, Poulsen, Klemm, Kröger & Schlierf, Establishing Superresolution Imaging for Protein in Diatom-Biosilica, 2016, Scientific Reports 6, 36824 

This project is part of the DFG Research Unit 2038: Nanomee