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Development of a wide-field TCSPC detector for FLIM TIRF microscopy of cell membrane receptors

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  • Full or part time
    Dr K Suhling
    Prof M Parsons
  • Application Deadline
    Applications accepted all year round

Project Description

Fluorescence microscopy techniques are powerful tools in the life sciences, because they are non-destructive, minimally invasive and can be applied to living cells and tissues. They can combine single-molecule sensitivity, molecular specificity, sub-cellular resolution and real-time data collection from live cells with negligible cytotoxicity, so that dynamics and function can be observed and quantified Fluorescence Lifetime Imaging (FLIM) in particular has emerged as a key technique to image the environment and interaction of specific dyes and proteins in living cells. FLIM can report on photophysical events that are difficult or impossible to observe by fluorescence intensity imaging, and FLIM is the best method to detect Förster Resonance Energy Transfer (FRET) to identify protein conformational changes or interactions. It is often implemented with beam scanning. However, some microscopy methods are best performed without beam scanning, employing wide-field camera-based detection instead, e.g. Total Internal Reflection Fluorescence (TIRF) microscopy. In collaboration with industrial partner Photek Ltd, we will develop a position-sensitive, picosecond resolution time-correlated single photon counting (TCSPC) detector with an electronic crossed delay line read-out for FLIM TIRF microscopy. This type of detector was used on NASA’s recent Horizon mission to study Pluto. The biological goal is to study the dynamics of receptor organisation at the surface of living human cells. Cell membrane imaging is performed with TIRF, a microscopy method using a camera, without beam scanning. It overcomes the current limitations of existing techniques and provides the high level of sensitivity, specificity and speed required - without beam scanning - to elucidate the control of receptor self-association at the membrane of living cells and how this is regulated by inflammatory insults. The work will be carried out in the Department of Physics and the Randall Division of Cell and Molecular Biophysics, in collaboration with Prof Maddy Parsons, and includes an industrial placement at detector manufacturer Photek Ltd.
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Funding Notes

Fully funded for 4 years.

Related Subjects

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