SSBII2

Scanning probe microscopy techniques are being increasingly applied to cell biological problems where novel applications have frequently reversed the drive for higher resolution and relied upon their precision in spatial and force control. We have designed and implemented a "life sciences platform" which integrates a TopoMetrix Explorer atomic force microscope (AFM) with a Nikon Diaphot 300 inverted optical microscope equipped with bright field video recording, fluorescent imaging, and laser scanning confocal microscopy via an integrated BioRad Radiance 2000 system. This has enabled us to image and analyse live cells in real time by AFM (e.g. topography, material properties, force distance analysis and binding site mapping) with simultaneous monitoring of cell morphology (e.g. time lapse recordings, reflection imaging, and fluorescence: GFP-labelled proteins, Fluo-3 intracellular calcium imaging, autofluorescence). Moreover, the ability of the AFM to act as a precise micro-manipulator under known applied forces has been utilised to deliver signals to cells and followed downstream phenotypic changes. This latter feature will have wide application to the analysis in real time of receptor-ligand interactions in live cell membranes and to investigate mechanically operated receptors and channels. Details of the experimental set-up will be presented and examples of the different analytical techniques