Confocal/Multiphoton Microscope Enhancements Provide Faster Scanning and Focus

Three enhancements to a laser scanning confocal and multiphoton microscope imaging system make it more powerful, provide new speed and sensitivity, and direct file access for ultra-large file acquisition and viewing.

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 (Center Valley, PA, USA) FV1000-MPE GaAsP detector is designed to provide much higher-sensitivity multiphoton 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.

With the new FluoView ASW 3.1 software, data sets of more than 150 gigabytes can be acquired in a single Z-stack. In addition, 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.

The Olympus technology also delivers better effective depth penetration, thanks to its optical design and pulse compression mechanism.

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