Nanobiotechnology Core Facility

Dissertation Thesis Topic: Membrane processes observed at nanoscale

Supervisor: Mgr. Jan Přibyl, Ph.D.

Annotation:

Membranes are the site of many processes in living organisms. Various aspects, including nanotoxicology, cell cytotoxicity, liposomes, extracellular vesicles (EVs) and nanomaterials will be studied, mainly using various microscopic and spectroscopic techniques, such as optical, fluorescence, AFM and Raman. The research should include characterisation of the synthetic bilayer, such as phase transitions and lipid domain formation, and further comparison with membrane studies in living cells. Changes in membrane properties under different physiological conditions will be studied. Fast processes can be observed using video rate AFM. This method should be carried out by the student in the laboratory with the newly installed instrument.

In addition, the effects of nanomaterial exposure on cell membranes will be studied using the above methods; changes in membrane morphology, roughness and mechanical properties in the presence of nanomaterials may clarify the correlation of nanotoxicity with structural changes. The work should also include the characterisation of nanoparticles whose shell is a phospholipid bilayer, i.e. a synthetic membrane. Such particles are liposomes and extracellular particles.

As part of his/her activities, the student should also participate in the activities of the core facility, but the focus of his/her activities will be his/her own scientific activity leading to a first author publication within the first 3 years of study.

Requirements on candidates:

In particular, the candidate should have an overview of the subject, a background in biochemistry or biology or related subjects, and above all a willingness to learn new things.

Keywords:

AFM, Raman, cell culture, nanoparticles, phospholipidic bilayer, nanotoxicology, membrane

Recommended literature:

1. Shaw, James E., Raquel F. Epand, Richard M. Epand, Zaiguo Li, Robert Bittman, and Christopher M. Yip. “Correlated Fluorescence-Atomic Force Microscopy of Membrane Domains: Structure of Fluorescence Probes Determines Lipid Localization.” Biophysical Journal 90, no. 6 (March 15, 2006): 2170–78. https://doi.org/10.1529/biophysj.105.073510.
2. Sadžak, Anja, Zlatko Brkljača, Mihael Eraković, Manfred Kriechbaum, Nadica Maltar-Strmečki, Jan Přibyl, and Suzana Šegota. “Puncturing Lipid Membranes: Onset of Pore Formation and the Role of Hydrogen Bonding in the Presence of Flavonoids.” Journal of Lipid Research 64, no. 10 (October 1, 2023). https://doi.org/10.1016/j.jlr.2023.100430.
3. Gyeong Lee, Hyo, Seokbeom Roh, Hyun Jung Kim, Seokho Kim, Yoochan Hong, Gyudo Lee, and Ok Hee Jeon. “Nanoscale Biophysical Properties of Small Extracellular Vesicles from Senescent Cells Using Atomic Force Microscopy, Surface Potential Microscopy, and Raman Spectroscopy.” Nanoscale Horizons 7, no. 12 (2022): 1488–1500. https://doi.org/10.1039/D2NH00220E.
4. Rikkert, L. G., P. Beekman, J. Caro, F. A. W. Coumans, A. Enciso-Martinez, G. Jenster, S. Le Gac, et al. “Cancer-ID: Toward Identification of Cancer by Tumor-Derived Extracellular Vesicles in Blood.” Frontiers in Oncology 10 (2020). https://www.frontiersin.org/journals/oncology/articles/10.3389/fonc.2020.00608.
5. LeClaire, Michael, James Gimzewski, and Shivani Sharma. “A Review of the Biomechanical Properties of Single Extracellular Vesicles.” Nano Select 2, no. 1 (2021): 1–15. https://doi.org/10.1002/nano.202000129.