Manager: David Jarrossay
This imaging facility (click to visit the web page) is central to most of the IRB research projects. The IRB has a state-of-the-art imaging facility (Flow cytometry and microscopy) headed by David Jarrossay who obtained a PhD degree at the University of Fribourg (CH).
The flow cytometry facility that is run by David Jarrossay who takes care of the cell sorting, maintenance of the equipment, instruction and advice to the new operators in addition to performing his own research.
Diego Morone, recruited in 2018, runs the microscopy facility. He is a microscopist and image analyst. He has experience in confocal microscopy, intravital microscopy and scientific image analysis. He teaches in the Microscopy Course organized by Prof. Thelen and aside offers a two-day internal course on Image Analysis with ImageJ.
The Flow Cytometry lab provides investigators with equipment and support for cell sorting (separation), acquisition, and analysis of flow cytometric data with a variety of state-of-art multicolor flow cytometry instruments.
- Cell sorting
Cell sorting is performed on a FACSAria III equipped with four lasers (488, 561, 640 and 405 nm excitation wavelengths) and 15 fluorescence channels detection. It can perform high speed sorting (up to 25,000 events/sec) with high-purity (up to 99%).
The lab also acquired a Sony SH800SFP cell sorter in a flow cabinet equipped with 4 lasers. Both sorters can perform cell disposition on plates.
- Benchtop analysers
The Flow cytometry lab is equipped with an advanced benchtop analyzer BD Fortessa equipped with four lasers (488, 561, 640 and 405 nm excitation wavelengths) and 16 fluorescence channels, one FACSCantoII (488, 640 and 405 nm lasers-eight colors) and FACSCanto I (488 and 640 nm lasers-six colors).
- High throughput screening
Beside both FACSCanto equipped with HTS for acquisition of 96 and 384 wells plates, the lab has also been equipped with an Intellicyt iQue (488 and 633 nm excitation-four colors) with automated platform (up to 45 plates per run) allowing high throughput screening for 96 or 384 well plates format.
The flow lab offers efficient support and high quality instruments. The staff provides cell sorting on BD FACSAria III, individual training on bench top analysers, maintenance of all instruments and assistance with experimental design, data analysis and troubleshooting. Three stations for data analysis (FlowJo software) are also available.
The main Microscopy Facility is equipped to perform most of the procedures for cell and tissue imaging (including FRET, FRAP, live cell imaging and intra-vital microscopy); it offers training on instruments, support for sample preparation, image analysis (Figure 3), deconvolution and 3D reconstruction; thanks to a wide range of software such as ImageJ, CellProfiler, RStudio, MetaMorph and MetaXpress (Molecular Devices), Matlab (MathWorks), Imaris (Bitplane).
The instrumentation includes wide-field fluorescence microscopes, high-content imaging system for image acquisition and analysis, confocal and multiphoton microscopes, comprising a surgical area for intra-vital microscopy:
- Confocal microscope. Leica TCS SP5, equipped with 4 high-sensitivity hybrid detectors (HyD Leica technology) and aberration-corrected objectives for high-resolution imaging (e.g. 100X 1.4 Oil)
- High-content screening system. The new Molecular Devices ImageXpress Micro 4 – installed at the beginning of 2018 – is a widefield automated microscope for the acquisition and analysis of plates and slides. It provides 5 acquisition channels, an incubator for timelapse imaging and a robot for automated acquisition of up to 45 plates.
- Wide-field microscopes.
- Nikon Eclipse E800 upright microscope;
- Nikon Eclipse TE300 inverted microscope, with incubator for live-cell experiments and Eppendorf FemtoJet microinjector;
- Zeiss Axiovert 200 inverted microscope, equipped with UV-corrected optics and FEI Live Acquisition software for calcium measurement experiments.
- Surgical microscopes.
- Olympus SZX10 equipped with a DP80 dual CCD color and monochrome camera for the imaging of tissues and fluorescence during the surgery.
- Leica M651 equipped with a MC170 HD color camera.
- Multiphoton Excitation microscopy system. LaVision BioTec TriM Scope, assembled system with upright and inverted microscopes, equipped with two tunable pulsed NIR lasers and OPO for multi-colour simultaneous imaging. Fluorescence is detected using new-generation hybrid detectors and infrared-corrected objectives, including a microendoscope