Background
Hepatocytes has elevated autophagy level and have been used to study various types of autophagy including micro autophagy, chaperone-mediated autophagy (CMA) and selective autophagy. We have identified a novel autophagy pathway with biochemical assays in rat hepatocytes, and the aim of the project is to perform imaging assays to identify further this novel auto-phagosome (Seglen, Engedal, Hu et al. publication in prep).
The imaging part of the project has several challenges. 1) Hepatocytes have strong autofluorescence which need to be examined and avoided while performing the immune-staining assay. 2) Hepatocytes are non-adhesive and the vesicles of interest are small, super resolution microscopy technique will be necessary.
During the project, the student will performe an imaging based screening study to identify other proteins that are present on the LDH positive autofagosomes. In addition the student will participate in projects where the same or similar compartments are to be defined in model cell lines.
A major aim for the student in this project is to get a broad basis of imaging techniques, image processing as expert in this field. In addition the student will be part of a team working on intracellular trafficking and will get a good background in standard molecular biology and cell biology techniques. Our imaging facility is part of Eurobioimaging as a node (https://www.eurobioimaging.eu/service), with many international activities.
Methods:
Molecular biology DNA techniques
Cell culture Techniques, expression of fluorescent tags in model cell lines
Super Resolution microscopy
Image Processing
Workplace and Environment:
Main part of the project is going to be conducted at the NorMIC Imaging Platform (NALMIN infrastructure Oslo node).