'White-light' flow cytometry

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Flow cytometry is a commonly used technique for analysing cells in which fluorescent molecules are used to optically tag cells that flow in a fluid stream through a laser beam. Detection and analysis of the fluorescence, generated as the cells traverse the beam, provides information on cell type and function. The technique allows extremely rapid analysis of large numbers of cells, as many as 1 million in a matter of minutes, and so can be used to detect extremely rare cells. For example fluorescent molecules can be introduced to cells within a blood sample and engineered, using anti-bodies, to selectively attach to any cancerous cells present. Flow analysis then identifies and counts these cells providing a measure of the number of diseased cells within the sample.This project will adapt a commercial flow cytometer, replacing the existing single colour laser with a multi-colour, 'white-light' laser. This provides laser light at wavelengths covering the whole of the visible spectrum and so can be used with any fluorescent molecule. Currently this choice is limited as each molecular species requires a specific wavelength of light to generate fluorescence. This unlimited availability of colour will allow multi-label experiments in which a large number of different fluorescent probes can be simultaneously introduced to the cell thus providing information on multiple interactions and processes within the cell. The modified cytometer will be used with lymphocytes (immune system cells) to gain a better understanding of childhood immunity in new born infants and its development during pregnancy.