Notre Dame Integrated Imaging Facility

The Nikon A1-R Confocal with Resonant Scanner, Spectral Detection, and Multi-Photon Modalities

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This powerful, fully-automated confocal imaging system captures high-quality confocal images of cells and molecular events at high speeds with enhanced sensitivity.  It is ideal for a broad range of users.  In the resonant scanner modality, high speed live cell observation (up to 420 fps) can be achieved during long term time-lapse imaging.  Working with a low pixel dwell time, one can image much faster with less photo-damage to the cells.  It can function as a spectral detector confocal with a 32-channel detector being simultaneously excited by up to four lasers, and wavelength resolution achieved by varying grating between 10nm/6nm/2.5nm.  This allows one to fine tune the acquisition parameters to the emission spectra of the chromophores within a sample, and enable accurate spectral unmixing for the separation of closely overlapping fluorescence spectra, and the elimination of auto-fluorescence.  This system can perform FRAP (Fluorescence Recovery After Photo-bleaching) experiments where one can simultaneously image the photo-activation of an area in a cell and capture the recovery or photo-conversion with no loss in temporal data.  In the Two-Photon or Multi-photon mode, the Nikon A1-R system enables imaging from deeper slices within specimens. It also uses a high speed scanner to take advantage of the benefits that come with two photon imaging – low photo-damage, deep imaging, and high speed (so you could image dynamic events deep in the sample in real time – not possible otherwise without the resonant scanner). 

The operating software is NIS Elements, a powerful software package that allows for integrated acquisition and analysis (from the 3D reconstruction, FRET analysis, object counting, and object tracking) that is intuitive and is actively being improved.  The base microscope is a Nikon Ti-E Inverted Microscope with Perfect Focus and motorized nosepiece and stage.  It works with the following lasers: 405, 488, 561, 640, and an IR laser for multi-photon imaging.  The following objective lenses available with this system are: 10x dry, 20x multi-medium (set up for oil), 40x oil, 60x oil, 100 x oil TIRF, 60x water (for multi-photon work), and 40x water (for multi-photon work).  It also has filter cubes and dichroic mirror sets to image in the fluorescent wavelengths of DAPI, FITC, TRITC, and Cy5.