High performance live cell imaging unit

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Recent advances in digital technology coupled with fluorescent microscopy allow the observation of events as they take place inside living cells. Traditionally, fluorescent microscopes consist of a source appropriate illumination that excites specimens and a light sensor that captures fluorescent emissions but its use has been limited by the lack of appropriate  technology to capture the dynamic interplay of cellular components in living cells.  However, during the last few years, fluorescent microscopes have been coupled with high-speed digital cameras allowing scientists to acquire fascinating data to test new hypotheses involving real-time cellular function. The equipment requested in this proposal encompasses such technology and will allow high-speed acquisition of fluorescent images from live cells without causing photodamage. The requested electron multiplier charge-coupled device camera (EMCCD), ImagEM (Hamamatsu), can acquire high resolution images at very low levels of light, making it ideal for high-performance live cell imaging. To control the camera and microscope and to process the acquired data, we are requesting the Volocity software package (Improvision), which provides a full suite of tools for 3D image acquisition, restoration, object measurement and tracking. Live cell imaging also requires the maintenance of optimal environmental conditions for the continued cell survival. Temperature and CO2 concentrations are fundamental variables for cell physiology, and must be kept constant in order to create proper experimental conditions at the microscope stage throughout the duration of the experiment. For this reason we are requesting the purchase of the Chalmide TC, a CO2/ Temperature controller device. This specific combination of equipment will comprise a unique Live Cell Imaging Unit (LCIU) that will be alternatively coupled to either an epifluorescent, a TIRF or a scanning confocal microscope already located within the Centre for Neurobiology of Stress facility at UTSC. An equivalent system does not currently exist at the UTSC campus and indeed this combination of apparatus is quite rare in Canada.