MICA: Imaging of cellular dynamics from single molecules to tissues

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Microscopy has been a key tool for giving insights into the structure and function of cells and tissues. The use of microscopy depends on achieving appropiate labelling/contrast of the molecules/structures of interest so that they can be visualised. In the past 20 years there have been enormous developments in microscopy. These have included the use of naturally glowing enzymes and proteins to visualise cellular processes. In addition, new "super-resolution" microscopes allow smaller structures to be visualised than ever before.This project will build upon our established track record in timelapse microscopy for the generation of cell movies of biological processes of interest. In the new Systems Microscopy Centre, we have state-of-the-art facilities and air-conditioned rooms specifically designed for microscopy experiments to generate movies of cells. We use the enzyme luciferase that makes fireflies glow to make cells glow (bioluminescence) when a chemical substrate is added. We also use a large number of different fluorescent proteins that glow in different colours when they are illuminated with different coloured light (fluorescence). The project is a close collaboration with the leading microscopy company Carl Zeiss, who will loan equipment which later will be purchased by the University once it has been optimised for the scientific applications.In this project we will combine bioluminescence and fluorescence so that different biological processes can be measured at the same time in single cells. This will allow us to watch cell signal timing at the same time as watching whether and when a gene becomes switched on to make a protein.We will also study a new super-resolution microscopy technique called SOFI where single molecules are located by watching the flickering in their light emission. This requires mathematical analysis to identify the exact localisation. Finally, we will use a microscopy method called "SPIM" that allows 3-dimensional biological structures to be studied. The principle is that a sheet of light is shone through the biological sample and then light emission by fluorescence is detected at 90 degrees to the plane of the light sheet. Often cells in the lab are cultured on glass which is clearly different to what normally happens in the body. Ultimately we hope to improve SPIM and to combine it with bioluminescence and the SOFI super-resolution approach.The project involves a large number of researchers with interests in a range of normal biomedical processes and diseases. Examples include inflammation, cancer, cardiovasular disease, and studies of how to grow new tissues from stem cells and the processes of normal embryo development. This project will act as a focus for future microscopy developments in Manchester University.