Label-free fluctuation-based superresolution microscopy with single-molecule sensitivity

Advisor: Marek Piliarik (IPE CAS, TPC DIGS-BB)

Funding: Fully funded

Website: https://cytoskeleton.eu/research/piliarik

Contact: piliarik@ufe.cz

While super-resolution microscopy has become an essential tool for understanding the structure and function of living systems, its fluorescence principle makes it difficult to image highly dynamic or complex systems. This blind spot in observing assembling protein structures hinders our understanding of some of the key processes that drive mechanisms in biological systems. One of the challenges inspiring our new technological developments is the earliest stages of tau aggregate formation. In neuronal cells, tau proteins regulate the access of other proteins to microtubules. Pathological self-assembly of tau is a hallmark of several neurodegenerative diseases, including Alzheimer's disease. To capture the mechanisms underlying biological systems in single-protein detail, the Ph.D. student will develop a new label-free method based on the direct detection of single-protein fluctuations using interferometric scattering microscopy (iSCAT). The development of the iSCAT method will focus on resolving microsecond fluctuations of single proteins, addressing details of the assembly of protein structures, such as tau proteins into fibrils and larger aggregates. The research aims to lay the foundations for a new era of high-speed, label-free super-resolution microscopy in biology and biophysics. The Ph.D. program is offered within the 1st Topical Partner Cluster (TPC) „Cytoskeletal Dynamics Across Scales: From molecular biophysics to organismal development“ associated with the Dresden International Graduate School for Biomedicine and Bioengineering (DIGS-BB).

Ph.D. student of the MFF UK under the supervision of the TPC member will be integrated into the DIGS-BB Ph.D. program including access to combined research infrastructure, techniques, training, lab rotations, and joint summer schools. DIGS-BB TPC provides expert mentoring and thesis supervision by each student’s Thesis Advisory Committee (TAC). Each TAC comprises three experts usually representing different disciplines and research institutions. The TAC will accompany each Ph.D. student throughout the thesis work for a maximum duration of 4 years. More information at https://www.digs-bb.de/tpc/cytoskeletal-dynamics-across-scales

[1] M. Vala, L. Bujak, A.G. Marin, K. Holanova, V. Henrichs, M. Braun, Z. Lansky, M. Piliarik, Nanoscopic Structural Fluctuations of Disassembling Microtubules Revealed by Label-Free Super-Resolution Microscopy, Small Methods (2021) 2000985

[2] H. Robert, K. Holanová, Ł. Bujak, M. Vala, V. Henrichs, Z. Lánský, M. Piliarik, Fast photothermal spatial light modulation for quantitative phase imaging at the nanoscale, Nature Communications 12 (2021) 2921

[3] M. Piliarik, V. Sandoghdar, Direct optical sensing of single unlabelled proteins and super-resolution imaging of their binding sites, Nature Communications 5 (2014) 4495

[4] M. Vala, M. Piliarik, Weighing single protein complexes on the go, Nature Methods 18 (2021) 1159–1160