Dr. Parazzoli’s imaging group, from left to right: Front - Simona Ronzoni, Anna Sciullo, Sara Barozzi and Erica Montani. Back - Dario Parazzoli, Massimiliano Garre’ and Amanda Oldani.
The IFOM-IEO Campus is a biomedical research centre, created by the joint efforts of the FIRC Institute of Molecular Oncology Foundation (IFOM) and the Department of Experimental Oncology of the European Institute of Oncology (IEO), which have expanded and integrated their research activities on a shared campus. The mission of the IFOM-IEO Campus is the development and application of genomics to research in basic and translational oncology. Its ultimate aim is to achieve a rapid transfer of the emerging concepts in the etiology and development of cancer to new strategies of treatment and prevention. Scientists at the IFOM-IEO Campus are investigating some of the most fundamental questions in cancer research, in fields ranging from molecular and cellular biology, genetics, functional genomics, proteomics, bioinformatics, systems biology and bioethics.
Research at the IFOM-IEO Campus is supported by a core technology facility that offers cutting-edge technologies in nanotechnology (DNA arrays), imaging, bioinformatics, robotics, proteomics, structural biology and animal models. This technology core is managed and coordinated by the "Cogentech" consortium.
The Imaging Centre of the IFOM-IEO Campus provides access to optical technology for biomedical researchers. Dr. Dario Parazzoli has been the director of this centre since the beginning of 2009. The major objectives of his unit are to set up, maintain and develop optical technologies to meet the needs of basic and advanced cancer research: flow cytometry and cell sorting, cell microinjection, wide field and confocal fluorescence microscopy, spectral imaging, deconvolution microscopy, total internal reflection fluorescence microscopy (TIRFM), two photon excitation microscopy (TPE), time-lapse microscopy, screening microscopy, advanced fluorescence based applications (FRAP, FRET and Photoactivation) and digital image analysis. The centre also provides sample acquisition and analysis in specific applications, on-site training and assistance to prepare researchers to the use of optical instrumentation. In addition, everyone working in the imaging centre performs independent and collaborative research for the development of advanced imaging-based technology.
Dr. Amanda Oldani is one of the members of the facility and her work is mainly dedicated to confocal microscopy. In particular, Amanda is responsible for the UltraVIEW VoX 3D Live Cell Imaging System.
Amanda says, “Working in this imaging facility is very stimulating because it gives you the opportunity to be in close contact with different research groups where you face varied and challenging requests. The investigation of biological processes connected with cancer development and diffusion cannot disregard the analysis of living cells and our researchers are more and more expressing the need for this kind of approach. The elected models are not always easy to handle and the use of the PerkinElmer UltraVIEW VoX spinning disk system allows us to get good results even with challenging samples. Its remarkable acquisition speed proved to be fundamental whenever extremely fast biological events are being investigated, such as retrograde actin flow (movie 1) or macrophage migration in living Medaka fishes (movie 2). The confocal capability of the system, linked to its high sensitivity and low phototoxicity, gave us the possibility to discriminate extremely dim signals above the background as in the case of protein recruitment on kinetochore in budding yeasts. Thanks to Volocity, not only can the spinning disk set up be easily tuned according to everyone’s need but also quantitative analysis and 3D-4D rendering can be effectively performed”.
Movie 1: The movie shows a HEK293 cell stably over-expressing proteins involved in adhesion and migration. After inducing cell spreading the actin backflow (visualized by Cherry-Lifeact) within the lamellipodium was recorded at a frame rate of 1 image per second. The time-lapse was performed using the UltraVIEW VoX spinning disk system, controlled by Volocity software. Movie courtesy of Carsten Schulte and Amanda Oldani.
Movie 2: The movie shows a z-projection of a 3D timelapse where wound-activated macrophages migrate within the Medaka (Oryzias latipes) tailfin. Membrane-tethered YFP-labeled macrophages (green) in the TG[FmpoP::mYFP] larva (Grabher et al., 2007), express transiently, and in a mosaic fashion, the nuclear marker, H2AmCherry (red). Images were acquired using the UltraVIEW VoX spinning disk system, controlled by Volocity software. Movie courtesy of Carolina Lage.
If you would like any further information about the imaging centre, please contact Dr. Dario Parazzoli.