Overview The laboratory for the Virus Imaging Core is currently located on the third floor of the Tarry Building on the Chicago Campus of Northwestern University. The laboratory will relocate to the state-of-the-art Lurie Biomedical Research Building in the fall of 2007. The Hope laboratory currently houses 3 DeltaVision RT deconvolution microscopes. Importantly, these microscopes are in a biocontained room located close to our HIV culture facility, allowing the safe observation of infectious HIV. Deconvolution microscopy uses computer processing to compensate for distortion caused by the optical path. Through the observation of fluorescent beads, an algorithm is developed which corrects the distortion caused by lenses and removes out-of-focus light163. The associated digital camera detects weak signals enabling great sensitivity and resolution, which is optimal for imaging viral particles in cells. Image capture with minimal illumination minimizes photobleaching and avoids phototoxicity, which can stress the cells during live cell observation. Two stand-alone workstations contain the complete Softworx software for deconvolutions and image processing, and off line image processing facilitates efficient instrument use. The systems are configured with filter sets allowing imaging of different types of fluorescent proteins including blue, cyan, green, yellow, and red fluorescent proteins. This permits simultaneous imaging of up to four different fluorescent proteins in live cells and 5 different fluors in fixed cell imaging. Importantly, two of these microscopes have environmental control chambers, including 5% CO2, allowing extended live cell timelapse microscopy. One of the microscopes with environmental control in equipped with a Photonics Mosaic digital diaphragm that allows precise sample illumination for photobleaching and photoactivation experiments. Additionally, this system has an ultrasensitive digital camera allowing maximal photon detection and signal amplification.
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