Live-cell hyperspectral microscopyGroup Leader Page
Project to be supervised jointly by Emmanuel Derivery and James Manton A typical human cell has around 10,000 different types of proteins, often interacting at specific locations for mere seconds or less. For example, the process of endocytosis by which cells internalise extracellular components involves ~60 proteins that act in a defined sequence at precise relative locations to each other. Our understanding of this and other biological processes would be greatly improved if we could directly visualise multiple proteins simultaneously with high temporal and spatial resolution in live cells. This project is aimed at realising this goal. You will develop a new, fast hyperspectral imaging system coupled to a high performance oblique plane light sheet microscope. This system will simultaneously visualise eight or more fluorescently-tagged molecular species at video rates, enabling live cell 3D hyperspectral imaging. After an initial year of development and testing, this system will be used to analyse the complex events of endosomal sorting and fission in live cells and ultimately organisms. In particular, you will make use of CRISPR lines expressing the proteins of interest fused to fluorescent tags at an endogenous level. You will be guided to develop skills in mechanical and optical engineering, cell culture and sample preparation, and image processing and data analysis. You will work synergistically with two teams: one expert in advanced microscopy development (James Manton) and the other in molecular dissection of trafficking and cytoskeleton processes (Emmanuel Derivery). By design, the module we propose to develop here can adapt to virtually any fluorescence microscopy technique, including super-resolution microscopy. Thus, we envisage the project expanding beyond endosomal trafficking to study the molecular interactome of other fundamental biological processes. This project would suit those with backgrounds in physics, biology, biophysics or bioengineering. A keen interest in both instrumentation development and solving fundamental biological questions is essential.
Emmanuel Derivery, Carla Sousa, Jérémie J. Gautier, Bérangère Lombard, Damarys Loew, Alexis Gautreau (2009) The Arp2/3 activator WASH controls the fission of endosomes through a large multiprotein complex Developmental Cell 17, 712–723. Emmanuel Derivery, Carole Seum, Alicia Daeden, Sylvain Loubéry, Laurent Holtzer, Frank Jülicher & Marcos Gonzalez-Gaitan (2015) Polarized endosome dynamics by spindle asymmetry during asymmetric cell division Nature 528, 280–285. Alex M. Valm, Sarah Cohen, Wesley R. Legant, Justin Melunis, Uri Hershberg, Eric Wait, Andrew R. Cohen, Michael W. Davidson, Eric Betzig & Jennifer Lippincott-Schwartz (2017) Applying systems-level spectral imaging and analysis to reveal the organelle interactome Nature 546, 162–167. Wiebke Jahr, Benjamin Schmid, Christopher Schmied, Florian O. Fahrbach & Jan Huisken (2015) Hyperspectral light sheet microscopy Nature Communications 6, 7990.