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BBSRC SWBio DTP Studentship: Developing a new bio-imaging tool for correlative light electron microscopy


Cardiff School of Biosciences

About the Project

Correlative Light Electron Microscopy (CLEM) is one of the most powerful imaging technologies as it combines the advantages of live cell imaging from light microscopy (LM) with the sub-nanometer spatial resolution of electron microscopy (EM). Using this technology, key biological questions have been answered [1]. CLEM is, however, seriously hampered by the availability of robust probes. It is highly questionable whether most bimodal probes, using a fluorophore (for LM) and an electron-dense gold nanoparticle (for EM) attached to the protein of interest, actually show the same protein pool. This is due to limitations such as photobleaching or quenching of the fluorophore, and/or detachment of the probes while trafficking inside the cell or after sample processing for EM. These are serious drawbacks that need to be addressed. To do so, the aim of this project is to develop a novel CLEM method where a gold nanoparticle (AuNP) is used as the same probe for both LM and EM. AuNP will be visualised in LM in living cells using a novel nonlinear optical microscopy technique developed at Cardiff University. The technique uses electronically-resonant Four Wave Mixing (FWM) and exploits the strong and photostable absorption and scattering of light of a AuNP at the localised surface plasmon resonance (LSPR). Using a combination of short optical pulses to generate and detect changes in the AuNP transmission or scattering at the LSPR, the technique is uniquely sensitive to single small AuNPs which are detected background free with high spatial resolution in 3D [2]. Notably, the technique is also uniquely sensitive to the AuNP shape, which opens up the prospect of probe multiplexing via shape recognition. In this project, spherical AuNPs and gold nanorods of various sizes will be conjugated to proteins of interest (for example the iron-binding protein transferrin) and internalised in mammalian cell lines. The aim will be to demonstrate the full CLEM workflow starting from imaging living cells with FWM (at Cardiff), to fixation, sectioning and correlative imaging of the same AuNP within the cell ultrastructure revealed by EM (at Bristol). The project will also explore the pioneering concept of assembling AuNPs directly inside living cells using proteins expressed in the cytoplasm which have the capability to bind metals and concentrate these to form electron dense particles. This could open the way to genetically tag cytoplasmic proteins with metallic NPs. [1] DOI: 10.1038/nature14503. [2] DOI: 10.1039/c9nr08512b


ELIGIBILITY
Applicants for a studentship must have obtained, or be about to obtain, a First or Upper Second Class UK Honours degree, or the equivalent qualifications gained outside the UK, in an appropriate area of science or technology. Applicants with a Lower Second Class degree will be considered if they also have a Master’s degree or have significant relevant non-academic experience.

In addition, due to the strong mathematical component of the taught course in the first year and the quantitative emphasis in our projects, a minimum of a grade B in A-level Maths or an equivalent qualification or experience is required.

Applicants must ensure they highlight their Maths background within their application and to upload any supporting evidence.

Funding Notes

A fully-funded four year SWBio DTP studentship will cover:
• a stipend* (at the standard UKRI rate; £15,285 per annum for 2020-2021)
• research and training costs
• tuition fees (at the standard UKRI rate)
• additional funds to support fieldwork, conferences and a 3-month placement

References

HOW TO APPLY
This studentship has a start date of October 2021. In order to be considered you must submit a formal application via Cardiff University’s online application service. (To access the application system, click the 'Visit Institution' button on this advert) 
There is a box at the top right of the page labelled ‘Apply’, please ensure you select the correct ‘Qualification’ (Doctor of Philosophy), the correct ‘Mode of Study’ (Full Time) and the correct ‘Start Date’ (October 2021). This will take you to the application portal. 
In order to be considered candidates must submit the following information: 
• Supporting statement 
• CV 
• Qualification certificates 
• Proof of English language (if applicable)
• In the research proposal section of the application, please specify the project title and supervisors of the project and copy the project description in the text box provided. In the funding section, select “I will be applying for a scholarship/grant” and specify advertised funding from BBSRC SWBio DTP. If you are applying for more than one Cardiff University project, please note this in the research proposal section as the form only allows you to enter one title.

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