Cell Imaging Facility

Making the discoveries that defeat cancer

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Cell Imaging Facility

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Cell Imaging Facility

Head: Dr Hugh Paterson

Introduction to Confocal Microscopy and Cell-Microinjection

The ability of cells to move is crucial for a range of biological processes; these include the development of organs, wound healing and inflammatory responses. However, the motility of cells can become deregulated in cancer leading to the spread of the cancer - a process called metastasis. Metastatic cancer is particularly dangerous and responsible for the majority of cancer deaths.

In order to study cell motility we combine confocal microscopy with time-lapse analysis to generate high-resolution videos of the dynamic behaviour of cells. Some of these videos can be viewed by following the links below.

The microinjection facility is based around a Zeiss/Eppendorf semi-automatic workstation, enabling experimental DNA expression vectors, recombinant proteins, or purified antibodies to be introduced directly into cells growing in culture. The imaging capability required for the analysis of such experiments has been significantly expanded to include two Bio-Rad MRC1024 confocal imaging systems. These permit simultaneous localisation of up to three intracellular proteins by immunofluorescence. In order to exploit the potential of Green Fluorescent Protein (GFP), conventional time-lapse video microscopy equipment has been upgraded by the installation of a cooled CCD-camera combined with digital image acquisition and processing. The high sensitivity of this system allows movement of specific GFP-tagged proteins expressed within living cells to be recorded over time and in response to extra-cellular stimuli.

Adherens Junctions

Epithelial cells use adherens junctions to stick together; these junctions often become disrupted as cancers progress. We have followed the behaviour of adherens junctions by following the distribution of a fluorescently tagged adherens junction component - GFP-alpha-catenin.

Time-lapse videos of GFP-alpha-catenin accompany our paper 'ROCK and Dia have opposing…' Sahai and Marshall: Nature Cell Biology 2002.

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hct116 cells expressing gfp-alpha-catenin

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hct116 cells expressing gfp-alpha-catenin and C3 toxin

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hct116 cells expressing gfp-alpha-catenin in calcium free media

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hct116 cells expressing gfp-alpha-catenin treated with nocodazole

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hct116 cells expressing gfp-alpha-catenin treated with cytochalasin D

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