| Supervisor Organisation | PhD Awarding Entity: | phd location |
|---|---|---|
University College Dublin | University College Dublin | University College Dublin, Ireland |
Research Focus
This project will employ soft X-ray technology and super-resolution microscopy to image disease proteins within the primary cilium cellular organelle. Current CLEM methods are cumbersome and technically demanding, and very few studies have probed how cilia disease proteins are arranged relative to underlying structure, especially in metazoans.
This project will investigate combined cryo-SXT, FIBSEM and cryo-SIM approaches to image fluorescent protein reporters in a leading animal model (C. elegans nematode) of human cilia disease, focussing on highly conserved pathways essential for cilium formation and function.
The project will develop new CLEM methods for possible roll out to other tissue and animal model contexts, and provide insight of disease mechanisms based on subciliary compartmentalisation of associated proteins.
Methodology
High pressure and plunge freezing methods will be implemented for preparing 20 micron thick slices of the nematode nose that contain primary cilia. Optimized techniques for correlative fluorescence/substructure imaging will be developed in collaboration with CLEXM partners; soft X-ray imaging will be performed with SiriusXT whose prototype benchtop microscope system is housed in UCD. Using optimised methodology, correlative imaging will be conducted on nematodes expressing endogenous (knock-in) fluorescence-tagged ciliary proteins.
Aim 1
Develop optimised methodologies for preparing and imaging nematode samples.
Aim 2
Determine nanoscale spatial distributions of fluorescence-tagged ciliary disease proteins, correlated with underlying subcellular structure
Aim 3
Assess how disease mutations impact correlated spatial distributions of fluorescence-tagged ciliary proteins.
Pictures Attached
The 1 mm long C. elegans animal model, and fluorescence and electron microscopy images of cilia in axial and radial orientations.
Role/Focus of PhD:
The PhD student will conduct all the experimentation and interpret the results. The student will also liase with collaborators and partake in a secondment to CLEXM partner laboratories. Successful applicants with have a BSc background in cell biology and have a passion for microscopy.
CLICK HERE TO GO TO THE WEBSITE PAGE
Application Deadline: 18th August 2023 (Closed)