A recently introduced multiphoton microscope system from Olympus allows researchers to achieve more sensitive, faster live cell imaging even deeper within tissue. The FluoView FV1200MPE multiphoton biological laser scanning imaging system is designed to deliver optimal live cell and tissue imaging. It offers increased laser selection, cooled high-sensitivity detector technology, increased scanhead light throughput and new software features designed for high speed functional measurements. The system can be built on the company’s innovative IX83 automated inverted microscope platform in addition to its BX61WI microscope for dipping-lens and live-animal applications. Olympus also introduced a confocal version of the FV1200 system this week.
Imaging living tissues demands the highest levels of sensitivity, and the FV1200MPE provides unparalleled sensitivity,along with new approaches for measuring rapidly changing fluorescent output, all while reducing laser power and destructive phototoxicity and photobleaching. The FV1200MPE is optimized for sensitivity, with increased excitation and emission transmission efficiencies. Its improved throughput enhances multiphoton excitation efficiency by more than 50%, maximizing the power available for the deepest imaging, uncaging or ablation needs. The system’s scanhead sets new performance standards with its breakthrough anti-corrosive silver-coated galvo mirrors that increase light throughput significantly over competitive instruments.
The new multiphoton system takes even greater advantage of enhanced transmission benefits by supporting the new Insight laser system from Newport Corporation for infrared (IR) imaging continuously from 690nm to 1300nm, enabling optimized excitation of longer-wavelength dyes as well as second- and third-harmonic-generation (SHG, THG) imaging. Dual simultaneousexcitation wavelength configurations are also available, either with the Insight Duo or with FV1200MPE TWIN configurations for simultaneous MPE imaging and stimulation, giving the FV1200MPE system one of the most robust and customizable sets of configurations available.
An optional high-sensitivity cooled gallium arsenide phosphide (GaAsP) detector unit also enhances sensitivity and minimizes electrical noise. When combined with new GaAsP detectors with high, 45% quantum efficiency, the unit provides some of the highest signal-to-noise performance possible. The combination of GaAsP and conventional PMTs in one system also allows users to choose either high-sensitivity detection or high-dynamic-range imaging, providing important flexibility for imaging live cells.
Additionally, the FV1200MPE is compatible with Olympus’ industry-leading range of specialty objectives, including the multi-award-winning SCALEVIEW objectives. Olympusmultiphoton systems are the only ones that boast compatibility with SCALEVIEW objectives, which extend the user’s ability to image at the deepest-ever levels. SCALEVIEW objectives are useful for large-scale connectomics analysis, in which researchers seek to understand connections across vast volumes of tissue athigh resolution. The 4mm and 8mm working distance, 25x MPE-dedicated SCALEVIEW objectives have already earned the R&D100, Edison, and Microscopy Today Innovation awards. The FV1200MPE also is compatible with dedicated 25x MPE objectives;Olympus microprobe “stick” objectives; and an expanded selection of live-cell-optimized silicone objectives including 30x, 60x and the brand new 40x high-numerical-aperture silicone immersion objectives.
The FV1200MPE is optimized for functional imaging. Its multipoint mapping advanced software (MMASW) can measure fluctuations in groups of cells at speeds up to 101 Hz in a full field, with sequential position data output up to 50,000 Hz, providing even fastermeasurements than are possible using raster-scanning resonance scanners. MMASW’s pseudo-heuristic point-scanning mode also offers significant improvements over raster scanning. It allows users to optimize their scan path as laser light moves from point to point, without any cropping; every scanned point also can be expanded to an array for larger-area stimulation or detection. The multipoint mode offers significant improvements over resonance scanners, swept field systems or acousto-optic deflectors (AODs) by retaining high integration time per position – where it is needed in a single point laser scan for greater multiphoton depth penetration and signal-to-noise performance.
Multichannel fluorescence measurements can be made precisely and accurately for such applications as calcium ion fluctuation, optogenetic experiments with channelrhodopsin and halorhodopsin, stem cell research, patch clamping/electrophysiology, and studies of functional output over cells andcell networks. The mapping feature allows pixel-skipping image generation in a field of fluctuating cells, along with automatic identification of regions of interest for further high-speed measurements. Users can take advantage of the ultra-high, physiologically relevant measurement speed while simultaneously stimulatingcells in complex multipoint patterns. The proprietary, highly efficient Olympus SIM galvo scanner is designed to operate simultaneously with the main scanner. Using tornado scan mode with the SIM scanner, and the optional Piezo-Z capability, the FV1200MPE provides optimized functional imaging deep within three-dimensional volumes for tissue or live-animal research.
The FV1200MPE takes advantage of all the enhanced touch-panel control, upgradeability, increased thermostability and flexibility of the new, state-of-the-art IX83microscope stand. The IX83’s U-shaped frame incorporates significant improvements in stability and rigidity, providing an ideal foundation for live cell experiments. The system is robust and highly expandable.
Filed Under: Drug Discovery
