| Imaging
Core Administrative | Protein Analytical | Imaging | Organic | Proteomics | Viral Production |
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DESCRIPTION OF THE IMAGING CORE Introduction The specific goal of the Imaging Core is to serve as a bridge to higher end imaging methodologies (i.e., confocal) by allowing researchers free access to microscopes and imaging software that will enable them to determine if more advanced analysis is needed or warranted. This Imagining facility strives to give investigators the opportunity to determine if their samples are worth a further investment of time and money to use higher end equipment on a fee-for-service basis. However, it should be noted that the Eclipse E600 microscope has been optimized for analysis of small cells such as yeast and platelets where high end microscopy (i.e., confocal) offers only limited benefit. The core also offers researchers an easy-access facility where simple analyses such as evaluation of transfection efficiencies can be monitored and documented. Finally access to the TIRF microscope offers a new horizon of technical capabilities that is only now becoming recognized by COBRE investigators. Facilities The facility has a Nikon Eclipse 600 epifluorescence microscope equipped with 10X, 20X, 40X, 60X, and 100X objectives that is capable of DIC and three-color analysis (Dapi, Texas Red, FITC). A Spot RT SE6 digital camera is attached to the microscope and images are captured and analyzed with MetaMorph imaging software run on a dedicated Dell computer. At the adjacent workstation is a Zeiss Axiovert 100 inverted epifluoresence microscope equipped with l0x, 32x, and 40x objectives that is capable of two color analysis (Texas Red, FITC). An Insight 11.2 Color Mosaic digital camera is attached to the microscope and images are captured using Spot imaging software run on a designated Apple computer. In the same room there is a Zeiss Axiovert 200M epifluorescence microscope with a motorized XYZ Stage (LudI) and a filterwheel housing filters for Dapi, FITC, TRITC and Cy5 as well as a DIC analyzer. The system is fully programmable for the XYZ coordinates, filterwheel positions and exposure time with a given set of filters. The microscope is equipped with an Orca ER camera (Hamamtsu, Inc.) which is capable of detection over a broad range of fluorochromes (far red to UV). The microscope system is controlled by a G4 Macintosh using Open Lab software. The system is designed for both live cells and fixed specimen imaging. Both the Bioptechs open and closed chamber heated stage as well as an objective heater are also available. Additional capabilities include time lapse and real time imaging of cellular events and deconvolution analysis of images. The Total Internal Reflection Fluorescence (TIRF) microscope, housed in room B074B, is built on an Olympus 1X71 inverted platform. An Andor iXON multichannel detector is used for image capture and image acquisition and processing software is run on a dedicated Dell computer. Each computer has full Internet access as well as access to laser printers and color laser printers. Training Opportunities Instruction on the proper use of the microscopes and interpretation of data are offered by Dr. Whiteheart on an individual basis as requested by investigators. Some remedial instruction is also available. Those individuals using the core receive instruction in microscope care and maintenance procedures. The core coordinates a series of instructional talks given by vendor representatives to better train users on the capabilities of the imaging software packages. Use by COBRE and Other Investigators
Project 1: Dr. Craig Vander Kooi
Project 2: Dr. Matthew Gentry
Project 3: Dr. Christian Paumi
Project 4: Dr. Francesc Marti
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