Over the last few years there have been incredible advances in microscopy and the associated software systems for image handling and processing. In parallel, there have been great developments in the use of fluorescent proteins, whose genes have been isolated from jellyfish and other marine organisms that glow in the dark, to label proteins within living cells. This allows for the first time, the location, movement, turnover and other behavioural aspects of human proteins to be seen in real time within living cells.

As a consequence, there is increasing demand for the study of live skin cells in culture in relation to our DEBRA-funded projects. For example, to observe the way normal and mutant keratins from EBS patients behave within cells being treated with our emerging gene therapy systems, such as ribozymes.

Existing available microscopes have a number of limitations. Firstly, they can only image static dead cells that have been chemically fixed and mounted on slides, rather than live

cells in culture. Secondly, these microscopes lack the capability to take optical sections i.e. an incremental series of images taken by focusing up and down through a given cell or tissue under computer control. Thirdly, the existing microscopes cannot be programmed to track cells at regular intervals as they migrate around culture dishes. Finally, our microscopes lack the capability to employ deconvolution software. The latter makes use of software modules developed for the Hubble space telescope and similar applications. This software takes a series of optical section images from a given target cell and processes these to remove essentially all background noise and construct a final image of incredible detail. This system is able to resolve structures within cells, particularly the cytoskeleton, with a level of detail unsurpassed by other types very high resolution microscopy.