A nano‐photocrafting technique for protein immobilization for nanoscopic cell studies

Activity: Talk or presentationOral presentation

Description

Nanoscale studies are essential for investigating intra- and extracellular behavior (e.g. cell activation and the formation of the cytoskeleton, uptake of EVs). To study these processes, a combination of nanocrafting to mimic physiological substrates and nanoscopic analysis for imaging and monitoring is required. Laser-Assisted Protein adsorption by Photobleaching (LAPAP) is a microfabrication technique to generate substrate-bound protein patterns. We are using a barcode system of oligonucleotides and the high binding affinity of biotin and streptavidin to bind biomolecules to the surface, which are presented to cells. The population of fluorophores in the triplet state and the lifetime of the triplet state are both crucial parameters for the efficiency of LAPAP. We established buffer systems that have photochemical effects on the fluorophores and therefore allow to control the number of bound proteins down to the single-molecule level. To analyze the bound proteins, we are using nanoscopic imaging, namely direct stochastic optical reconstruction microscopy (dSTORM) and stepwise photobleaching for single-molecule analysis. Besides new possibilities for cell experiments on surface-bound proteins, our buffers have a high potential to be used in water-based photoresists to further increase the resolution in nanolithography.
Period14 Sep 2021
Event titleMICROSCOPY 2021: CSMS Conference
Event typeConference
LocationČeské Budějovice, Czech RepublicShow on map