Spinning Disk Confocal Microscope for Intravital Imaging

The intra-vital microscopy system is based on the Yokogawa Corp.'s CSX M1-L scanning disk upgraded to 5000 rpm (1000 scan/sec). The system allows high speed image acquisition at high resolution and with low photo-toxicity. This system has an AOTF controlled laser array providing 405, 488, 514, 568, 647 nm laser lines. A custom computer drives a stage with piezo 300 µmZ axis control for acquisition of multi-color Z stacks. The system includes emission band pass or long pass filters (Chroma): 460/50, 525/50, 605/52, 700/75. These filters allow the detection of DAPI-CFP/GFP, FITC/mCherry/Cy5. For image acquisition, the system includes a Hamamatsu EMCCD 512×512 driven by Micro-manager 1.4 open source software.