Hollas, Alexis C.

Holmes-Smith, A. Sheila

Hungerford, Graham

McLoskey, David

The use of fluorescence techniques to study biological media is well established. In particular fluorescence lifetime imaging microscopy (FLIM) is a powerful tool for studying kinetic processes within the cell membrane. The study of environmental influence on intercellular properties, such as viscosity, is of importance. Yeast is a good model system in which to observe this. Its reproductive rate and size, allow dyes to be taken up and changes influenced by the growth media explored via microscopy. In this work we employ the fluorescent probes, Dimethylaminostyrylmethylpyridiniumiodine (DASPMI) and 2-chloro-4-(2,3-dihydro-3-methyl-(benzo-1,3-thiazol-2-yl)-methylidene)-1-phenylquinolinium iodide (FUN-1) to stain Saccharomyces cerevisiae. DASPMI is a viscosity sensitive mitochondrial stain while FUN-1 is a viability sensitive stain, therefore, changes in viscosity and intracellular activity will be reflected in the fluorescent properties of these dyes. However, one of the problems with live cell imaging using FLIM is that the cell can move during the time it takes to make the measurement; this therefore impacts on the quality of the image. Here this is overcome by immobilising the yeast cells within a biocompatible medium based on a polysaccharide, which allows cell growth, but minimises cell diffusion. In this work we present fluorescence lifetime imaging of immobilised yeast cells, demonstrating the viability of the cells and explore the cell growth rate under different environmental conditions

Fluorescence lifetime imaging of immobilised saccharomyces cerevisiae

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