Imaging and Cytometry Laboratory, Department of Biology, University of York.

Imaging and Cytometry Laboratory, Department of Biology, University of York.

The Imaging and Cytometry Laboratory houses a comprehensive suite of microscopy and cytometry equipment and provides research support and training to a wide range of people from both the university and external organisations. The Imaging and Cytometry Laboratory was founded as part of the Technology Facility, which itself is comprised of six laboratories within the Department of Biology at the University of York, providing a unique, multi-discipline research support and training unit. Dr Peter O’Toole came in as head of the Imaging and Cytometry Laboratory which was initially funded through part of a BBSRC JIF award, and the laboratory has since grown to include over £3 million worth of equipment. This has been achieved through further BBSRC, SRIF and commercial income.

Peter’s unit provides a facility committed to bringing state-of-the-art technology to researchers with varying levels of expertise. A significant effort is devoted to training both internal and external (academic and commercial) scientists and the team provides fully assisted technical support for infrequent users and an excellent equipment resource for experienced users. Working closely with the equipment manufacturers, the team is also involved in various aspects of technology development.

Peter gained his PhD in the Cell Biophysics Laboratory at the University of Essex and has been involved in many aspects of fluorescence imaging. Since arriving at York, he has initiated several courses to teach all aspects of confocal microscopy to a diverse range of participants. He is a member of the executive council of the Royal Microscopical Society for which he organizes the Light Microscopy School, confocal course and the practical flow cyctometry course. For more information about the Technology Facility and the courses available please visit their website.

The laboratory houses one spinning disk microscope, two laser scanning confocal microscopes and one multiphoton microscope with three non-descanned detectors for increased sensitivity at greater depth. These are used extensively for many sample types, including live cell studies, tissue sections and morphological studies of other substrates, enabling co-localisation with multiple fluorescent probes and advanced techniques such as FRET, FRAP, FLIM and Spectral Unmixing. The laboratory also has a number of wide-field fluorescent microscopes and a laser capture microdissection microscope to enable live, cultured cells or cells of interest from tissue sections to be identified, excised and catapulted. These rare cells can be re-cultured or exploited, for example through PCR analysis. All of the systems are linked to digital cameras for high quality image capture and subsequent image processing. The lab has an off-line analysis suite that includes an Improvision License Server with five Volocity Licences. The server allows researchers anywhere in the Department of Biology to perform 3D and 4D visualization and analysis back in their own lab. Alongside the light microscopy equipment, the laboratory also houses a number of electron microscope and flow cytometers, allowing seamless integration between these two complementary approaches.

One of the major users of Peter’s unit is Dr Mark Coles and his research group from the Immunology and Infection Unit at the University. Research in the Coles lab utilises 4D multiphoton imaging, 3D whole mount confocal microscopy, gene expression analysis and tissue engineering to study the development and function of primary and secondary immunological organs. Mark’s laboratory uses Volocity for Microsoft Windows XP Professional x64 Edition for imaging 3D and 4D data sets using an 8 processor/16GB workstation. “Volocity 4.3 x64 allows us to see cell movement in 4D in real time, this used to take a significant amount of time before and is a major time saving to members of the lab.” Recent research in the lab has used lineage tracing tools to analyze the embryonic origin of stromal cells in lymphoid organs (Journal of Immunology, 180(5):3183-9, 2008).

To learn more about the Imaging and Cytometry Laboratory, please visit the facility website.

T cell Zone stromal cell network (Red) in peripheral LNs (Imaged by Dr. Priyanka Narang)