LAboratoire de Spectrochimie Infrarouge et Raman – UMR 8516
random_1
  • Français
  • English
  • Spanish

Descriptif du sujet

Improved methods for spatial-temporal fluorescence imaging of cell activity
Encadrement : C. Ruckebusch, (. , U. Lille), . Peter, D. (KU L. ,
Sujet : 

The PhD projects aim to develop a new methodology to extract superresolution spatial-temporal information on cell activity in response to some external stress. This will provide new insights into how biological systems are structured at the nanoscale, and how this structures provides e.g. signaling specificity, using the latest methodologies in imaging and data analysis.

One of the most important issues in biology is understanding signaling activities in living cells. How does the cell transfer specific signals through a complex interaction network in order to trigger an action in response to a stressing event? To achieve this transduction, the cell must regulate its response by managing the spatial temporal organization of its constituents. This means that a given molecule not only has to be present in the cell but has to be there in the right place at a particular time. As a consequence, simply visualizing the spatial distribution of molecules is not sufficient. Recent advances in (chemically smart) fluorescent labels have made possible to visualize and quantify the cellular activation dynamics in living cells. Information about when and where activity arises is assessed following how the fluorescence emission of the probe changes conditional on some aspects of the environment. One approach consists of mapping Forster Resonant Energy Transfer activity at an interesting spatial and temporal resolution. However, the construction of diffraction unlimited and high-sensitivity superresolution FRET activity maps requires acquiring and interpreted multicolor donor acceptor data, as well as developing specific data and image analysis methods to handle stationary signals, deal with strong photobleaching and low S/N to noise, or to unravel signal multiplexing.

REFERENCES

-Super-resolution imaging goes fast and deep, S. Duwe, P. Dedecker, Nat. Methods 14:1041 2017

-Improved superresolution microscopy imaging by sparse deconvolution with an interframe penalty, S. Hugelier, P.H.C. Eilers, O. Devos, C. Ruckebusch, J. Chemometr. 31 1– 9 2017

-Correcting for photodestruction in super-resolution optical fluctuation imaging Y. Peeters, W. Vandenberg, S. Duwé, A. Bouwens, T. Lukeš, C. Ruckebusch, T. Lasser, P. Dedecker, Sci. Rep. 7 10470-1 -10470-10 2017

-Sparse deconvolution of high-density super-resolution images, S. Hugelier, J. De Rooi, R. Bernex, S. Duwé, O. Devos, M. Sliwa, P. Dedecker, P.H.C. Eilers, C. Ruckebusch, Sci. Rep. 6 21413-1 – 21413-10 2016