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Fluorescent Nanosensors for Biological Measurement


   School of Pharmacy

  , Prof Jon Aylott  Friday, November 05, 2021  Funded PhD Project (UK Students Only)

Nottingham United Kingdom Analytical Chemistry Bioinformatics Biophysics Nanotechnology Optical Physics

About the Project

AIM: This exciting EPSRC Funded project will be to develop and apply fluorescent nanosensors for biological measurement. This will produce an advanced platform that will optimise disease prediction, diagnosis and intervention, through developments of new understating of biological microenvironments.

BACKGROUND: Fluorescent nanosensors are inert, versatile biosensors, that can be used to make important measurements of key molecules and ions in microenvironments at the physics of life interface and are ideal for real-time measurements of dynamic processes [1].

At the University of Nottingham, fluorescent nanosensors have been developed to quantify pH, [2] molecular oxygen [3] and temperature [4] in complex model systems. Of these sensors, the pH-sensitive fluorescent nanosensors have gathered the greatest momentum. They have been evaluated and validated in a range of complex and diverse microenvironments and demonstrated their immense potential by mapping the acidification in nematode model organisms (Caenorhabditis elegans [5] & Pristionchus pacificus [6]), elucidation of subcellular fermentation pathways in Saccharomyces cerevisiae [7], determined the intracellular processing of foreign material in human mesenchymal stem cells (hMSCs) [8] & characterised the evolution of acid by-products during bacterial biofilm growth [9].

This project will continue the application of pH-sensitive fluorescent nanosensors to diverse biological environments, which include fruit flies (Drosophila melanogaster), Thale cress (Arabidopsis thaliana) and CHO/Heck 293 cell culture for bioprocessing. As well as develop new biosensors for key biological molecules and ions.

 IMPACT: The new understanding gained from innovative analytical tools developed as part of this project will advance micro-environmental insights in biological systems, currently not possible with conventional techniques, such as probes and free fluorophores. This will ultimately lead to the optimisation of disease prediction, diagnosis and intervention through improvements in understanding of ion and molecule flux in biological microenvironments.

This project will provide diverse training opportunities for the PhD candidate in fluorescence, microscopy, spectroscopy, analytical chemistry, particle characterisation and model system culture. With support from the experienced supervisory team, the research conducted will pave-the-way towards establishing new methods towards the understanding the complex and dynamic functions of ions and molecules in diverse biological systems. This data obtained will be used to generate high impact publications and opportunities to disseminate research at international conferences.


Funding Notes

This position is open to national and international candidates.
• Candidates must possess or expect to obtain, > 2:1 degree in Physics, Chemistry, Biology, Pharmacy or related scientific discipline, which include Bioinformatics and Mathematics
• The project is suitable for candidates with a scientific background who would like to develop their experimental analytical skills.
• A background in bioinformatics and mathematics is desirable.

References

[1] S. Ahmed, V.M. Chauhan, A.M. Ghaemmaghami, J.W. Aylott, New generation of bioreactors that advance extracellular matrix modelling and tissue engineering, Biotechnology Letters, 41 (2019) 1-25.
[2] V.M. Chauhan, G.R. Burnett, J.W. Aylott, Dual-fluorophore ratiometric pH nanosensor with tuneable pK(a) and extended dynamic range, Analyst, 136 (2011) 1799-1801.
[3] V.M. Chauhan, F. Giuntini, J.W. Aylott, Quadruple labelled dual oxygen and pH-sensitive ratiometric nanosensors, Sensing and bio-sensing research, 8 (2016) 36-42.
[4] V.M. Chauhan, R.H. Hopper, S.Z. Ali, E.M. King, F. Udrea, C.H. Oxley, J.W. Aylott, Thermo-optical characterization of fluorescent rhodamine B based temperature-sensitive nanosensors using a CMOS MEMS micro-hotplate, Sensors and Actuators B-Chemical, 192 (2014) 126-133.
[5] V.M. Chauhan, G. Orsi, A. Brown, D.I. Pritchard, J.W. Aylott, Mapping the Pharyngeal and Intestinal pH of Caenorhabditis elegans and Real-Time Luminal pH Oscillations Using Extended Dynamic Range pH-Sensitive Nanosensors, Acs Nano, 7 (2013) 5577-5587.
[6] J.W. Lightfoot, V.M. Chauhan, J.W. Aylott, C. Rödelsperger, Comparative transcriptomics of the nematode gut identifies global shifts in feeding mode and pathogen susceptibility, BMC research notes, 9 (2016) 142.
[7] M.M. Elsutohy, V.M. Chauhan, R. Markus, M.A. Kyyaly, S.J.B. Tendler, J.W. Aylott, Real-time measurement of the intracellular pH of yeast cells during glucose metabolism using ratiometric fluorescent nanosensors, Nanoscale, 9 (2017) 5904-5911.
[8] R.P. Harrison, V.M. Chauhan, D. Onion, J.W. Aylott, V. Sottile, Intracellular processing of silica-coated superparamagnetic iron nanoparticles in human mesenchymal stem cells, Rsc Advances, 9 (2019) 3176-3184.
[9] B. Blunk, M. Perkins, V.M. Chauhan, J.W. Aylott, K.R. Hardie, Fluorescent Nanosensors Reveal Dynamic pH Gradients During Biofilm Formation, bioRxiv, (2020) 2020.2007.2031.230474.

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