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The Light Microscopy Unit at the PKZI
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The Cell Biology Unit of UMC-Mainz apart from the research unit encompasses the core facility of advanced light microscopy. Prof. Dr. Gregory Harms heads the advanced light microscopy platform which will not only cater the needs of colleagues interested in advanced light microscopy techniques but also aims to further develop and equip the infrastructure in the next years. In partnership with Leica, the facility will serve as a "Reference Center" and thus will gain access to the new developments in microscopy, image analysis tools and other unique techniques.
In addition to the microscopes present in the Core Facility, there are an SP8 CLSM and a DMi8 time lapse microscope available in the AG Rajalingam. If you are interested to use the machines, please contact either Hermann Götz (hgoetz(a)uni-mainz.de), Christiane Schönfeld (schoenfeld(a)uni-mainz.de) or Gregory Harms (greharms@uni-mainz.de) for further information.
Leica TCS SP8 DIVE System
The Leica TCS SP 8 DIVE is an upright fixed stage microscope based on a DM6 CFS stand. The motorized xy stage is meant for in-vivo as well as in-vitro observation of thick tissue materials or small animal applications. A specialized waterimmersion objective with motorized corection is mounted (IRAPO 25x/1.00 NA Water). An IR Laser (Mai Tai HP DeepSee) with tuneable 690 - 1090 nm wavelength is used for the multiphoton exitation and a 3-channel reflected light detection system RLD-NDD (1PMT, 2 HyD) for the image aquisition. For conventional confocal image aquisition a 2-channel PMT detection and 2 lasers (488 nm, 552 nm) are available. For high speed image aquisition a 8 kHz tandem scanner is used to overcome challenges such as small movements at live animal observation.
Leica TCS SP8
The Leica TCS SP8 is an inverse confocal fluorescence microscope based on an DMi8 CEL advanced stand. A stage top incubation system allows live cell imaging with multicolor fluorescence. Five lasers are available for excitation (405, 488, 514, 552, 638 nm). The fluorescence emission can be continuously adjusted between 400 - 800 nm and is detected by 2 PMTs and 1 HyD. A HPZ 640 premium computer with CUDA grahic card and the Leica HyVolution software (Huygens Deconvolution) permits online confocal superresolution. Mosaic scanning of large objects is possible due to the motorized xy-table and the Navigator Software tool. Following objectives are available: 10x/0.30 dry, 20x/0.75 dry CS2, 63x/1.30 GLYC CORR CS2, 63x/1.40 Oil CS2.
Leica TIRF
The Leica DMi8 S TIRF is an inverted widefield microscope for live cell imaging. It is equipped with a complete incubation housing system for temperature and CO2 controlling. An external light source (EL 6000) for conventional fluorescence microscopy and phase contrast methods is mounted. It is further equipped with a TIRF (= total internal reflection) module for infinity port, which is ideal for the imaging of samples located close to the cover glass/water interface like membrane receptors. In TIRF mode four lasers are available for excitation (405, 488, 561, 638 nm) and a specialized objective (100x/1.47 TIRF Oil). The fluorescence emission is detected by a very fast and sensitive Leica DFC9000 GTC sCMOS camera. For the conventional mode following objectives are also available: 5x/0.12 PH0, 20x/0.40 CORR PH1, 40x/0.60 CORR PH2.
Image acquistion and data processing
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Leica LAS X 3D Analysis: A HPZ 440 Computer with offline LAS-X software licenses is available for measurement, 3D visualisation, 2D- and 3D image processing and analysis and the Huygens Deconvolution Software.
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Bitplane Imaris Server: For image processing and analysis the Bitplane Imaris 9.1 Software is available on an optimized PC computer system. The package consists of one set of licenses for Imaris including each of the following modules: MeasurementPro, ImarisTrackLineage, ImarisColoc, ImarisXT, ImarisCell, FilamentTracer, ImarisVatage and ImarisBatch. It is ideally suited for researchers who need the entire functionallity of Imaris on a single dedicated Workstation.
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