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BP: Fachverband Biologische Physik

BP 2: Bioimaging and Spectroscopy I

BP 2.5: Vortrag

Montag, 20. März 2017, 10:45–11:00, HÜL 386

Exploring protein diffusion landscapes in living embryos with SPIM-FCS — •Philipp Struntz, Dirk Hofmann, and Matthias Weiss — University of Bayreuth, Experimental Physics I, Germany

Macromolecule diffusion in the complex and dynamic environment of living organisms often features spatial variations that report on the cells' secret life, e.g. during embryogenesis. To explore these spatial heterogeneities one needs to quantify the local diffusion characteristics in extended regions of the sample in a multiplexed fashion. To obtain diffusion maps with high spatiotemporal resolution we have combined single plane illumination microscopy (SPIM) and fluorescence corre- lation spectroscopy (FCS). By refining a custom-made SPIM setup that was originally designed for long-term in-vivo imaging of early embryos of the small nematode Caenorhabditis elegans [1], we were able to acquire pixel-wise FCS curves on spatially extended regions within the embryo. We demonstrate the capabilities of SPIM-FCS by determining the diffusion maps of the peripheral membrane protein PLC1δ1 in the cytoplasm and on the plasma membrane during early stages of embryogenesis [2]. In a next step, we have focused on time-resolved diffusion maps of the protein PIE-1, for which we see the formation of a mobility gradient along the anterior-posterior axis before the first, asymmetric cell division. Our data hence show that SPIM-FCS can be used to explore intracellular transport phenomena even in fragile developmental model organisms.

[1] R. Fickentscher, P. Struntz & M. Weiss, PRL 117, (2016).

[2] P. Struntz & M. Weiss, J. Phys. D 49, 044002 (2016).