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Multiphoton Microscope System

Fri, 02/28/2014 - 2:23pm

The Olympus FluoView FVMPE-RS, a dedicated multiphoton microscope system, enables high-precision, ultra-fast scanning and stimulation, allowing researchers to see deep within specimens, take measurements at the highest speeds and capture images. The FVMPE-RS is designed for electrophysiology and optogenetics studies. It is also a good match for applications such as high-speed calcium and in vivo imaging, peristalsis and blood flow studies, mosaic imaging, connectomics and functional brain imaging, stem cell research and any field that requires precise colocalization, uncaging, simultaneous imaging/stimulation, extensive real-time signal processing, or multipoint mapping. Its design offers ready adaptability for researchers who design their own custom-built optical delivery as well. The FVMPE-RS captures 438 frames per second (fps) at 512x32. It also captures full-frame, 512x512 images at 30fps without any reduction of the field of view, a critical feature for many functional imaging studies. Its scanner unit combines a newly designed resonant scanner with a galvanometer scanner to provide both speed and definition. The resonant scanner offers an exclusive non-linear sampling method for smooth imaging without intensity variation or image distortion, and greatly minimizes line jitter at any zoom factor. An optional third galvanometer scanner is also available for simultaneous imaging with 3D stimulation or uncaging in a given optical section. The system’s multipoint mapping capability allows ultra-high-speed stimulation along with measurement of rapid fluctuations in groups of cells for functional imaging studie, 3D intensity measurement or mapping with high signal-to-noise performance.

The FVMPE-RS provides multicolor, multiphoton excitation and imaging with a choice of lasers, along with an industry-first four-axis auto-alignment capability for precise colocalization and coalignment without pixel shift. With its multiple-laser-line performance, researchers can image with two colors at optimal wavelength excitation simultaneously while stimulating the sample with visible light, or image with one infrared (IR) laser while simultaneously stimulating with IR and/or visible light. The system delivers sharp images with clear separation of fluorescence proteins for analysis.

The high-sensitivity system offers excellent throughput from 400nm to 1600nm. New 1600 Coating technology provides broadband transmission without sacrificing the ability to stimulate at lower wavelengths (such as 405nm for glutamate uncaging or 458nm for channelrhodopsin stimulation). The new coating allows the system to support the latest InSight lasers from Spectra-Physics, providing researchers with the ability to image near-IR dyes such as Cy5 and Cy7. The Olympus line of 25x multiphoton optimized objectives also feature this new coating, and offer working distances of 2, 4 or 8mm to deliver super-deep imaging. Light collection efficiency is further improved by the FVMPE-RS system’s optimized light path. In Deep Focus Mode, researchers can adjust laser properties precisely to account for tissue scattering, helping optimize deep-tissue imaging by achieving signal improvements of 20 percent or more over comparable systems. An optional high-sensitivity cooled gallium arsenide phosphide (GaAsP) detector unit further enhances sensitivity while minimizing electrical noise, making the most out of the system’s high-speed capabilities. 

The system’s precision timing allows for microsecond repeatability and control of multiple imaging and stimulation protocols, as well as millisecond repeatability over days of time-lapse imaging. Complex multi-position imaging or optogenetic stimulation protocols can be accomplished using the advanced stage control and sequence manager.  The new system’s multichannel analog box for triggering, synchronization and signal input provides additional support for advanced electrophysiology studies and other applications requiring external device triggering.  The advanced optional nosepiece piezo drive provides greater rigidity and faster z-plane stabilization to ensure accurate and repeatable high-speed focus performance.

Olympus America, Inc., 800-446-5967, www.olympusamerica.com/FVMPE-RS

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