Olympus Confocal/Multiphoton Microscope Enhancements Provide Faster Scanning and Focus, Higher Sensitivity, and Ability to Acquire Ultra-Large Images

CENTER VALLEY, Pa. – Three enhancements to the well-respected and widely used Olympus FluoView® FV1000 laser scanning confocal and multiphoton microscope research imaging system make it more powerful than ever, providing new speed, sensitivity and direct file access for ultra-large file acquisition and viewing. Researchers will benefit from a gallium arsenide phosphide (GaAsP) detector, updated ASW v.3.1 software and a new multipoint mapping/scanning and piezo focus system. All provide important advances in performance and usability.

One enhancement is designed for neuroscientists and cell biologists who must visualize very weakly fluorescing specimens, highly photosensitive samples or very thick specimens requiring the highest sensitivity. The Olympus FV1000-MPE GaAsP detector is designed to provide much higher-sensitivity mulitphoton detection for these samples. The unit offers higher signal-to-noise performance and more than double the quantum efficiency of standard photo-multiplier tubes (PMTs), for greater imaging depth and sensitivity. It couples two standard PMTs and two additional GaAsP channels for use when the highest sensitivity is required, providing flexibility. The cooled GaAsP detector also has a protection circuit to ensure the safety and integrity of the system.

Olympus ASW v.3.1 software helps researchers acquire large Z-stacks of image data and view them easily. Until now, it has been difficult to acquire and view data at Nyquist sampling rate over a large Z depth; this is due to the large file sizes created. But with the new FluoView ASW 3.1 software, data sets of more than 150 gigabytes can be acquired in a single Z-stack. Even more importantly, image files can be accessed directly from the hard disk rather than having to be loaded into RAM, thanks to improved memory handling. With the ASW v.3.1 software’s enhanced graphical user interface (GUI), users can easily acquire well beyond Nyquist rate and open their images directly within the software, without having to transfer their image data to other programs, making data review easy. The direct-access feature is designed to work with the FV1000’s automated stage, making acquisitions of hundreds of gigabytes possible.

For physiologists and others who need to do fast measurements at multiple points or tissue depths (such as calcium signaling in cell networks), the Olympus high-speed piezo-Z focus scanning mechanism provides the speed necessary for fast data acquisition over multiple Z-axis locations. The technology offers better signal-to-noise ratios (S:N) than other scanner technologies because most other commercial technologies use dwell times that are too short to maintain high S:N. In addition, the beam positioning used by many systems may not be as accurate as that of the Olympus system; even averaging the scans captured using these other technologies may not fully make up for the difference. The Olympus technology also delivers better effective depth penetration, thanks to its optical design and pulse compression mechanism.

When coupled with the ASW v.3.1 software’s optional multipoint mapping advanced software (MMASW) module, the piezo-Z system provides a powerful tool for researchers doing electrophysiology or high-speed measurements such as calcium signaling. The software is designed to provide the speed researchers need and the data they want at a range of depths. The multipoint mapping software provides the equivalent of 101-frame-per-second (fps) performance in a given Z-plane by using advanced pseudo-heuristic technology. This means that researchers can look at the travel time between points and optimize the path of travel to maximize the number of positions; they also can spread each position’s measurements out over a larger area. At each sampling cycle, high-speed fluctuations within each position are read out every 20 microseconds for a user-defined interval, even when measuring at multiple depths within tissue.

By using a combination of pseudo-heuristic scanning and piezo-Z, researchers using the Olympus system can conduct an experiment where they might select and measure 10 points per plane in 10 different Z-axis planes in less than one second, with a remarkable 50,000-Hz data set generated at each point. Using the piezo-Z system, typical bi-directional Z-scanning speeds are improved as well, allowing multi-depth frame scans to be captured at video rate. Transistor-transistor logic (TTL) synchronization allows the piezo system and MMASW software to interface with electrophysiology equipment and other devices.

“The best multiphoton system just got even better,” said Brendan Brinkman, product manager for Olympus America Inc. “Researchers have long relied on FV1000 confocal and multiphoton microscope systems because of their performance and unsurpassed optics, and users now have the additional benefits of faster, more sensitive, more flexible and more easily accessed data than ever before. The piezo-Z and pseudo-heuristic scanning bring new power to functional imaging, and when the software and GaAsP detector are coupled with our recently announced SCALEVIEW 25x, 4mm working distance multiphoton imaging objective, the implications for large-data applications such as the connectome are profound.”

For more information on the piezo, GaAsP or ASW v.3.1 enhancements to the FluoView confocal/multiphoton microscope system, visit www.olympusamerica.com/fv1000accessories or contact Brendan Brinkman, Olympus America Inc., phone 424-298-7402 or email [email protected].

About Olympus America Inc., Scientific Equipment Group

Olympus America Scientific Equipment Group provides innovative microscope imaging solutions for researchers, doctors, clinicians and educators. Olympus microscope systems offer unsurpassed optics, superior construction and system versatility to meet the ever-changing needs of microscopists, paving the way for future advances in life science.

About Olympus

Olympus is a precision technology leader, designing and delivering innovative solutions in its core business areas: Medical and Surgical Products, Life Science Imaging Systems, Industrial Measurement and Imaging Instruments and Cameras and Audio Products. Olympus works collaboratively with its customers and affiliates worldwide to leverage R&D investment in precision technology and manufacturing processes across diverse business lines. These include:

• Gastrointestinal endoscopes, accessories, and minimally invasive surgical products;

• Advanced research, clinical and educational microscopes and research and educational digital imaging systems;

• Industrial research, engineering, test, inspection and measuring instruments; and

• Digital cameras and voice recorders.

Olympus serves the healthcare field with integrated product solutions and financial, educational and consulting services that help customers to efficiently, reliably and more easily achieve exceptional results. Olympus develops breakthrough technologies with revolutionary product design and functionality for the consumer and professional photography markets, and also is the leader in gastrointestinal endoscopy and clinical and educational microscopes. For more information, visit www.olympusamerica.com.

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