The Olympus FluoViewTM FV500 is a point-scanning, point-detection, confocal laser scanning microscope designed for biological research applications. Excellent resolution, efficiency of excitation, intuitive user interface and affordability are key characteristics of the Olympus FV500. A fully automated confocal system, the FV500 may be configured with ultraviolet through infrared lasers and permits simultaneous collection of up to 5 detection channels. The FV500 scanning unit can be coupled to either the IX2 inverted or the BX2 upright research microscope platforms.
Olympus FV500 Confocal Microscope Features
Confocal microscopy can improve conventional fluorescence images by recording fluorescence generated from the focal plane within the sample, while rejecting all other light coming from above or below the focal plane. The efficient point-scan/pinhole-detection confocal optics of the FluoViewTM systems virtually eliminate out of focus light to produce high-contrast images with superb resolution. The features listed below are available on all Olympus FV500 microscope system configurations.
Versatility in Scanning Modes
The FluoViewTM confocal microscopes are equipped with several efficient scanning modes, including point, line, free line, and rectangle, which are especially suited for many time-lapse applications.
Laser Systems Offer Large Selection of Wavelengths
Laser systems designed for the Olympus FV500 confocal microscopes have a broad spectrum of available wavelengths with intensities selectable by individual neutral density filters or the acousto-optic tunable filter (AOTF) controller for simultaneous or automated-sequential collection of multi-channel images. A choice of the 440-nanometer diode laser and the 442-nanometer helium-cadmium laser is available for fluorescent protein (cyan and yellow, CFP/YFP) resonance energy transfer applications, providing optimal excitation of CFP and spectral separation from the YFP. The shutters and light intensity can be controlled via the system computer.
The 405-nanometer diode laser, a low cost alternative to water-cooled ultraviolet lasers, can be used for most ultraviolet imaging applications, including nuclear imaging with DAPI. Unique laser modulation of the 405-nanometer diode laser permits high-speed control of the light intensity, which can be important for photoactivation and photobleaching studies. The exceptional light stability of the 405-nanometer diode laser is also important for time-lapse studies of living cells.
In the FV500, as many as four high-sensitivity photomultiplier tubes can be incorporated directly within the confocal fluorescence emission light path for high sensitivity detection of the fluorescence signal. A fifth photomultiplier detector, dedicated for transmitted light imaging, may be used for the simultaneous detection of high-resolution brightfield or DIC images with which the confocal fluorescence images may be overlaid.
Depending on configuration, up to five of the detection channels may be imaged simultaneously. Images may be scanned in any pixel array size up to 2048 x 2048 pixels, with each channel digitized to 4096 gray levels (12-bit), permitting observation of fine image detail. Images may also be acquired in an automated sequential mode in order to reduce spectral crosstalk between channels for multi-color images.
The FV500 microscopes incorporate independent confocal apertures in front of each photomultiplier fluorescence detector. Each confocal aperture is continuously variable and software-controlled for an optimal match of each aperture diameter to the objective magnification, numerical aperture, and emission wavelength settings.
Automated software control of the FluoViewTM confocal microscope laser excitation, emission dichroics, and barrier filters provide a simplified setup of the system's hardware. Simply select the combination of fluorochromes to be imaged and the software automatically configures the complete imaging pathway for each channel to be recorded.
Flexibility in the selection of the microscope base, laser lines, and optical filters permits researchers to customize their Olympus confocal system for a broad range of research applications.