The Fluoview 200 system is designed to acquire high spatial resolution images of fluorescently labeled materials and for analysis of these images. The confocal principle utilizes a pinhole (confocal aperture) to eliminate out-of-focus light from fluorescently labeled specimens (i.e., provide “optical sectioning”). Lasers provide intense, point illumination that is scanned over the preparation and the fluorescence at each point is quantified and used to construct a representation of the object brightness (an image). This method provides high resolution for the x and y planes as well as vertically (z plane).
The Fluoview is based on an inverted microscope to facilitate the study of live materials in special chambers or, to a limited extent, in culture plastics. For practical reasons, your specimens should be visible through a #1 coverslip to utilize the high resolution available. This may require growing your cells on coverslips or fitting a coverslip window into culture plastics to aid observation (if interested, ask how to do this).
The Fluoview uses two lasers to produce 3 excitation lines (488, blue; 543, green; and 633, red). This allows excitation of green (FITC, GFP, BODIPY, etc), red (rhodamine, Texas red, phycoerythrin, Cy3, propidium, etc), and far-red dyes (Cy5). You will not be able to excite blue dyes (DAPI, Hoechst, even CFP is poor). In addition, the system provides differential interference contrast (DIC) images as well as reflectance scan modes. There are two detectors that can operate simultaneously. Z sectioning is automated and has a step size resolution of 0.1 µm. Images are 12-bit (4096 levels of gray/intensity) and range from 256×256 to 1024×1024 pixels (picture elements). Image acquisition time ranges form 1sec
The Fluoview is inexpensive as confocal systems go, but it is far from cheap! Use of the confocal system requires in-person training to make sure the system remains available to Users. Laser safety must be followed! There is a hourly fee for using the Fluoview, and Users will be billed to recover costs incurred in maintaining the system. The fee schedule is currently $17.00/h for NPP / Cancer Center folks, $20 for other UC staf.
Get pdf Fluoview 200 Laser Scanning Confocal Microscope Operation Manual
