Fabian Wehnekamp will be visiting from the Ludwing Maximillians University, Munich via the International Physics of Living Systems Student Research Network (iPoLS).
The talk will be held Thursday, Feb. 12, at 2 pm, location TBA.
Orbital tracking: Performing 3D real-time particle localization in vitro, cells and live tissue
Life depends on the shuffling, transport and relocation of proteins on the cellular level. For a better understanding of these processes, the ability to follow and track particles in 3D with high spatial and temporal resolution is crucial. To meet this challenge, we have developed a home-build Orbital Tracking microscope with simultaneous widefield observation capabilities with which we are able to follow a nanoparticle in real-time with nm precision in a complex 3D environment. I will discuss the development, theory and applications of this technique ranging from in vitro systems to zebrafish (danio rerio).
In the second part of this talk, I will focus on mitochondrial dynamics in the sensory neurons of zebrafish. A malfunction in mitochondrial dynamics is one possible cause for neurodegenerative diseases (tauopathies). To follow the trajectory of individual mitochondria in rohon-beard sensory neurons, we coexpressed photoactivatable GFP and TagRFP by coinjecting neuron specific driver constructs (Isl2b) with separate UAS responder constructs into fertilized single-cell zebrafish eggs. Zebrafish expressing the markers are measured individually three days post fertilization. By photoactivating individual mitochondria, we are able to obtain single-mitochondria trajectories with a traveled distance of more than 100μm with nm precision. Due to our high spatial and temporal resolution, we can identify several different dynamic populations involved in mitochondrial transport. The environmental information gives insight into the interaction between stationary and moving mitochondria. By combining the results from the fast and precise orbital-tracking microscope with the widefield data, we obtain an in vivo overview regarding the dynamic processes in rohon-beard sensory neurons that can be used for further disease-related investigations.