Dr. Barbara Vertel
The big one
the second one
The Werner Straus Live Cell Imaging Laboratory is a core facility in the Department of Cell Biology and Anatomy that offers access to state-of-the-art, high-speed multidimensional image analysis specifically designed for living cells. The laboratory provides expertise in light imaging techniques that include 4D data acquisition, FRAP (Fluorescence Recovery After Photobleaching), FLIP (Fluorescence Loss In Photobleaching) and FRET (Fluorescence Resonance Energy Transfer). These experimental approaches are widely used to study molecular dynamics in live cells, highlighted by the use of green fluorescent protein and its variants to visualize protein trafficking. The facility houses a Nikon Imaging System and a Zeiss LSM 510 Confocal System, which are available on a fee-for-use basis (see below). A separate off-line PC workstation with Metamorph and AutoQuant Deconvolution software and the Zeiss LSM software is available for post-acquisition image analysis and data processing. Specific arrangements must be made with the Lab Directors before projects are initiated. In-house training will be given to all new users and consultation regarding imaging aspects of your research project will be provided. Investigators will supply their own samples and materials.
The Nikon High-Speed Multidimensional Image Analysis System is a wide-field system specifically designed for visualizing live cells. Based on a Nikon TE2000U Inverted Microscope, the system includes a Roper/Photometrics Cascade 16 Bit high-speed CCD camera, filter wheels and shutter, x-y motorized stage and z-focus control. It is optimized for simultaneous time-lapse acquisition of images at multiple wavelengths and z-focus, at multiple stage positions. Filters and/or filter cubes are available for a variety of fluorochromes, including DAPI/Hoechst, GFP/FITC, TRITC/Texas Red/Cy3 and Cy5. Plan Apochromat objective lenses that are available include 10x/0.45 DIC, 20x/0.75 Phase/DIC, 40x/0.95 Phase/DIC, 60x/1.4 oil Phase/DIC and 100x/1.45 oil TIRF DIC. The custom designed microscope environmental chamber provides stable, long-term temperature control and the capability for CO2 regulation. An external MicroPoint laser ablation system can support FRAP and FLIP analysis in combination with multidimensional imaging.
The Zeiss LSM 510 Confocal Microscope System is available for live cell imaging and many photobleaching techniques. It is equipped with multiple excitation laser lines at 405nm, 488nm, 543nm and 633nm. The three separate detection channels are capable of detecting many standard fluorophores covering the visible light spectrum including: DAPI, CFP, GFP, YFP, Fluorescein, Rhodamine, Texas Red, Cherry, RFP, Cy5 and several of the Alexa Fluor dyes. This is an inverted microscope (Axiovert 100M) with an automated stage and universal stage insert that can accommodate several types of specimen slides and live chambers. The available objective lenses include: Plan Neo 10x/0.3, Plan Apo 20x/0.8, Plan Apochromat 63x/1.4 Oil DIC and Plan Apochromat 100x/1.4 Oil DIC. The system can be used to image single, double and triple-labeled specimens, analyze and quantify colocalization of fluorophores and conduct a live time-lapse series. In addition, it is designed for the photobleaching techniques including fluorescence recovery after photobleaching (FRAP), inverse FRAP, fluorescence loss in photobleaching (FLIP) and fluorescence resonance energy transfer (FRET).
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