Notre Dame Integrated Imaging Facility

The Andor/Nikon Spinning Disk Confocal Microscope

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Andor Technology and Nikon Instruments worked to produce this Spinning Disk, live cell confocal microscope.  This system is designed to image thin samples with low emission fluorescence as well as capture cellular events with minimal photo-damage to the cells.  It does this by combining Andor’s operating software iQ2, EMCCD cameras, and solid-state laser with Yokogawa’s CSU-X1 confocal scanner and Nikon’s Ti-E Inverted Microscope with Perfect Focus and motorized nosepiece and stage.  This provides a framework for laser-illuminated microscopy solutions.  In addition to spinning disk confocal, other modalities that can be performed with this system include: TIRFM (Total Internal Reflection Fluorescence Microscopy observes the properties of an induced evanescent wave at the interface or the contact area between a specimen and a glass coverslip or tissue culture container), FRET (Fluorescence Resonance Energy Transfer microscopy techniques show protein interactions that occur between two molecules positioned within several nanometers of each other), and FRAP (Fluorescence Recovery After Photo-bleaching, the recovery process of fluorescence over time can be observed to analyze molecule diffusion rate).  The imaging software Andor iQ2 is designed with multi-dimensional, live-cell, and time-lapse image acquisition, processing and analysis, and therefore has many powerful features not found in conventional image analysis products.  Also installed is Molecular Devices Meta Imaging Series MetaMorph software that may be used to operate the system for users that are more familiar with it than Andor’s IQ2 software.  The objectives lenses for this instrument are a 20x dry, 40x oil, 60x oil TIRF, and 100x oil TIRF.  Also available upon request is a long working distance (2.8mm to 3.6mm) 40x dry lens. The following laser modules are found in the solid-state laser combiner: 405nm (50 mW), 488nm (25 mW), 561nm (25 mW), 640nm (40 mW).  These excitation lines work with the emission fluorescence filter cubes and custom dichroic mirror sets to detect fluorescence in the following wavelengths: DAPI, FITC, TRITC, and Cy5.