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An evaluation of liquid-liquid phase separation in disease cells and in vitro using fluorescence lifetime imaging and image correlation spectroscopy


   Faculty of Biology, Medicine and Health

  ,  Applications accepted all year round  Self-Funded PhD Students Only

About the Project

Liquid–liquid phase separation is a mechanism used by eukaryotic cells to organize their intracellular content; it has linked to cell malignancy and neurodegenerative diseases. Inside cells, are formed membrane-less organelles such as nucleoli, centrosomes, Cajal bodies, and stress granules, all based on liquid–liquid phase separation (LLPS). LLPS depends on parameters such as mixture composition, structure, and temperature; these membrane-less organelles may represent “active liquids” in a metastable state and appears to use ATP to control their fluidity; however the formation, existence of LLPS and the relation to ATP remains poorly understood. Recent studies have suggested that fluorescence lifetime imaging and an image correlation spectroscopy variant have shown potential to monitor the presence of LLPS in cells. Here we aim at evaluating the formation of membrane-less vesicle/droplets in vitro first then in cells varying parameters such as composition, temperature to understand whether these microscopic techniques can provide reliable information. These will be compared against techniques such as turbidity, static light scattering for in vitro experiments.

Entry Requirements

Candidates are expected to hold a minimum upper second class honours degree (or equivalent) in a physical chemistry, physics, biophysics, chemical engineering or any related subjects. Candidates with experience in any of the aforementioned techniques or with an interest in liquid-liquid phase separation are encouraged to apply.

How To Apply

For information on how to apply for this project, please visit the Faculty of Biology, Medicine and Health Doctoral Academy website (https://www.bmh.manchester.ac.uk/study/research/apply/). Informal enquiries may be made directly to the primary supervisor. On the online application form select the appropriate subject title.

For international students, we also offer a unique 4 year PhD programme that gives you the opportunity to undertake an accredited Teaching Certificate whilst carrying out an independent research project across a range of biological, medical and health sciences.

Equality, Diversity and Inclusion

Equality, diversity and inclusion is fundamental to the success of The University of Manchester, and is at the heart of all of our activities. The full Equality, diversity and inclusion statement can be found on the website https://www.bmh.manchester.ac.uk/study/research/apply/equality-diversity-inclusion/”


Funding Notes

Applications are invited from self-funded students. This project has a Band 2 fee. Details of our different fee bands can be found on our website (View Website).

References

Kanaan et al,. “liquid-liquid phase separation induces pathogenic tau conformations in vitro” Nature Communications (2020), 11: 2809
Pliss and Prasad “high resolution mapping of subcellular refractive index by fluorescence lifetime imaging: a next frontier in quantitative cell science?”, Methods and applications in fluorescence (2020), 8, 032001.
Wohl et al., “Fast and synchronized fluctuations of cortical actin negatively correlate with nucleoli liquid-liquid phase separation in T cells”, Eur. J Biophysics (2020), 49, 409-423.
JW Bye and RA Curtis “Controlling phase separation of lysozymes with polyvalent anions”, J Phys Chem B (2019), 123(3), 593-605

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