Hybrid Techniques

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The ScanIC head may be mounted on an inverted optical microscope, for visual selection of target areas and to correlate optical and SICM imaging. Other systems may be linked in, providing powerful extensions to a range of investigative techniques.

Patch-clamp

Replacing micromanipulators with a ScanIC offers radical improvement to patch-clamp productivity and effectiveness. The ScanIC's vertical pipette is normal to the cell surface, enabling the patching of cells unattainable until now due to their low profile - e.g. neurons or spermatozoa. Also, use of the ScanIC may obviate the need for an optical microscope.

Using the patching pipette itself as the scanning probe, a cluster of cells may be imaged by the ScanIC and a target cell selected. That cell may be scanned at higher resolution to select a suitable 'landing site'. The ScanIC can then steer the probe to its target and to land it vertically on the surface.

Throughout this process, the tip is protected from inadvertent contact with the cell surface or with floating debris by the ScanIC's control system.

Surface confocal microscopy

Mounted on an inverted confocal microscope, the ScanIC probe tip and laser focus may be locked together, while scanning is achieved by movement of the sample. The confocal laser then tracks the pipette tip as it follows the sample surface contours. As a result, fluorescent emissions across the sample surface may be mapped in a single pass, eliminating degradation by photo-bleaching. As well as saving the time required for multiple 'slices', this procedure enables the derivation of meaningful quantitative data that is comparable across the entire surface. Importantly, by setting the laser focus to a fixed distance below the pipette tip, fluorescent emissions may be mapped, in a single pass, at a given depth below the surface.

Scanning near-field optical microscopy

In fluorescence-mode SNOM, accurate and stable control of the tip-to-surface distance is essential. SICM is an ideal control system for SNOM of biological samples, employing a pipette whose tip outer surface has been coated with aluminium by evaporation, coupling a laser light source to the pipette through a multi-mode optical fibre and collecting scattered light through an inverted microscope configuration.

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