Super-resolution imaging of human peroxisomes - Master student project 2022

Peroxisomes are key organelles involved in a large variety of metabolic processes, which depend on organism, tissue and developmental stage. Pex3 is an important peroxisomal membrane protein that is proposed to play a role in peroxisome biogenesis, with mutations leading to severe disorders. It is proposed to play a role in multiple peroxisome-related processes, all requiring different binding partners in yeast. This makes the interpretation of mutant phenotypes very complex. Most likely, human Pex3 is also involved in additional functions, but these have not been identified yet. Partially because the small size of mammalian peroxisomes (~200nm) hinders unveiling peroxisomal structures by conventional light microscopy.

Therefore, stimulated emission depletion (STED) nanoscopy will be used to study the (co-)localization of human Pex3 on peroxisomes in detail. This super-resolution technique enables fast (live-cell) imaging beyond the diffraction limit of light (30-40nm in cells), without further data processing.

In this project, we will mainly work with Human Embryonic Kidney (HEK) cell lines. The techniques will involve cell culturing and labeling of the cells with STED-suitable dyes or antibodies. Next to this, we will optimize imaging settings on the STED microscope. Hereby, we will use fixed as well as living cells. After that, the image visualization in ImageJ and data analysis will also be part of this project, to experience a complete cycle from beginning to end.