Real-Time Microscopy System (Inverted Olympus IX 70) with image analysis and electrophysiology capabilities
The Real-Time Microscopy System is built around an Olympus IX-70 inverted microscope. It is connected to three light sources, including a Sutter DG4 excitation monochrometer, and a Hamamatsu OrcaER CCD camera interfaced with a Sutter W-10 emission filter wheel. The Simple PCI software controls these illumination delivery and emission collection devices, along with driving the microscope's light path shutters and automated focus. The workstation's foundation is a vibration isolation table and is shrouded by a darkened, faraday cage. The microscope has been adapted with variable stage warming and environmental gas controls. This system has provided users with high-speed/short-term imaging (e.g., calcium imaging) and slow acquisition/long-term imaging (e.g., GFP expression) platforms. Electrophysiological studies are conducted using a mobile electrophysiological workstation from the neurobiology laboratory of Mark Zoran, including patch-clamp amplifiers and a digital data acquisition system.
The Real-Time Microscopy System has been used extensively for studies of calcium imaging in neurons and glia. Studies of intracellular calcium in diencephalic and retinal glial cell cultures have demonstrated the utility of these cultures for investigation of the role of melatonin, its receptors, and its signal transduction mechanisms in the regulation of intercellular calcium waves (V. Cassone Lab).
Imaging studies of gap junctional coupling have demonstrated that melatonin affects changes in intercellular coupling that are inconsistent with a major role for gap junctions in the propagation of calcium waves.
The Real-Time Microscopy System has also been extensively used for studies of GFP expression/imaging in cultured suprachiasmatic nucleus (SCN) neurons (D. Earnest Lab).
