The mission of the Microscopy and Cytometry Facility (MCF) is to provide expertise and support in experimental work involving advanced microscopy of biological specimens, and cell sorting. The current services that are provided by the facility can be divided into three groups.
The cell sorting service is provided using a Becton Dickinson FACSAria II and Beckman Coulter CytoFLEX SRT cell sorters. FACSAria II is equipped with three lasers (violet, blue, and red) and nine fluorescence detectors. CytoFLEX SRT is equipped with four lasers (violet, blue, yellow-green and red) and fifteen fluorescence detectors. Up to four populations can be sorted simultaneously. Cell sorting is offered as a full service for occasional customers and users and as equipment access for researchers who are experienced in flow cytometry.
The facility provides access to a broad range of fluorescence light microscopes. Most of our microscopes allow optical sectioning, such as confocal (point-scanning or spinning-disk), two-photon, lightsheet, and total internal reflection fluorescence (TIRF), to facilitate high-contrast fluorescence imaging. The facility’s newest acquisition is Opera Phenix, a high-content screening system from PerkinElmer for the large-scale imaging of cells (e.g., in RNAi-based microscopy screens) in widefield or confocal mode. Our equipment also includes a Zeiss LSM800 confocal microscope with a high-resolution Airyscan detector, a Zeiss LSM710 NLO dual confocal/multiphoton microscope for the live imaging of cells and tissues, an Andor Revolutions XD system for real-time spinning-disk confocal microscopy and TIRF imaging, a Zeiss Lightsheet Z.1 single-plane illumination microscope for the imaging of fluorescently labeled zebrafish larvae, an Olympus CellR/ScanR imaging station for intracellular calcium measurements, and a Nikon 80i Eclipse microscope with a scanning stage for the mosaic imaging of histochemically or fluorescently stained tissue sections. Two- and three-dimensional image analysis is possible using dedicated software, such as Imaris (Bitplane) and Harmony (PerkinElmer). Full imaging services are also possible on the most sophisticated microscopy platforms.
The electron microscopy service offers analyses of cells, tissues, and virus particles with a FEI Tecnai T12 transmission electron microscope. For the conventional transmission electron microscopy of cells and tissue samples, we use a Leica EM tissue processor. This enables resin processing under constant temperature while avoiding exposure to toxic substances. After saturation with resin, tissue and cell specimens are pre-trimmed with a Leica EM TRIM2, which prepares for the next step of processing. Samples are then cut for semi- and ultra-thin sections using a Leica EM UC7 ultramicrotome, and then sections are placed on electron microscopy grids. Material that is prepared this way can be imaged with our electron microscope.
The MCF operates in either full-service mode or access mode, depending on equipment, application, and the customer. In the latter mode, our staff offers initial training for users and assistance with experimental design, data analysis, and final data interpretation. The MCF is open to IIMCB researchers and external customers from academia and industry.
The detailed list of equipment with description and contact names is available under the “Equipment“ tab in the “Core Facilities” menu.