Olympus FluoView 500 Laser Scanning Confocal Microscope LSCM
The FV500 is controlled by a 2.4 Ghz personal computer under Windows XP and is capable of imaging 5 separate channels simultaneously (4 fluorescence + 1 transmitted light photomultiplier detectors) offering highly efficient, maximum emission sensitivity and the ability to record scanned images in 12 bits or 4096 gray levels, thus allowing quantitative linear measurement of fluorescence within regions of low contrast as well as very high contrast. Users are able to image a wide variety of fluorophores with laser excitation that includes Blue Violet (405nm) Multi-Line Argon Blue (458,488,515nm), Helium Neon Green (543nm) and Helium Neon Red (633nm) for standard Blue, Green, Red and Far-Red fluorochromes. The FV500’s acoustical optical tuning filter (AOTF) and adjustable scan speeds provides for minimal specimen fading, sequential scanning for reduced fluorescence cross talk, multiple regions of excitation, high resolution imaging (up to 2048 x 2048 pixels) of fixed or static samples, and rapid recording of kinetic events.
Optical sections in the z plane can be collected using a stop motor attached to the fine focus control of the microscope and driven by Fluoview software. The system is also equipped with Differential Interference Contrast (DIC) objectives and condensers and has the ability to capture transmitted light images with a highly sensitive photomultiplier (PMT) transmission detector.
Images are saved to local secondary 250 GB hard drive and then transferred to a 2 TB mirrored RAID-1 network-attached storage (NAS) device that is accessible from core workstations or individual laboratory computers through the UM medical campus 4 Gbps network. Integral Fluoview software allows for analysis of saved images in 2 dimensions (e.g. brightness vs time); confocal images obtained in a z-series can be volume rendered and analyzed in 3 dimensions. Fluoview software saves images in a non-proprietary format to facilitate the use of the raw data files in 3rd party image analysis or visualization software such as ImageJ, MetaMorph, AutoQuant, Volocity and Imaris.
Strengths of the Olympus FluoView 500 Laser Scanning Confocal Microscope
- Great work-horse system for routine 1, 2 or 3 color (blue, green, red or green, red, far red) imaging of fluorescent signals in fixed cells or tissue.
- Filter based system to direct wavelengths to the PMT detectors. In some instances signals are stronger compared to spectral detectors.
- Cy-5 filter set for visualizing far red signals in the oculars/eyepieces before imaging with the laser.
Confocal images taken in the MICPC