Bioortogonální značení povrchových receptorů na nefixovaných lymfocytech

Paldusová, Kateřina

The surface of cells displays high heterogeneity on chemical and geometrical levels. To understand the function of cells, we need to pay attention to the morphological features formed at the plasma membrane. To study cell surface with molecular specificity, there are plenty of imaging methods starting with the conventional wide-field microscopy through confocal microscopy, ending with super-resolution fluorescence microscopies and electron microscopies. Super-resolution microscopy studies conducted on the fixed cells provide detailed steady-state data about the cell surface nanoscopic organisation and distribution of molecules at the morphological structures. However, since cells are parts of living organisms and constantly change their properties in time and space, the information about dynamics of cellular structures and motility of molecules remains hidden when using this approach. Live-cell compatible methods are required to study dynamic changes of molecules at the single-molecule level. In this study we are focusing on the distribution and dynamics of molecules CD2 and CD4 expressed on the surface of non-stimulated T cells. The main aim of this thesis was to develop a novel method for live-cell imaging and single-molecule tracking of membrane-bound proteins in 3D and at nanoscale. With such a...

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