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Multifocal Multiphoton Microscopy Using a Novel Field of View Zoom Scanning Protocol

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We present a novel field of view (FOV) zoom scanning multifocal multiphoton microscopy (MMM) system that is based on a microlens array for producing discrete excitation spot array, and the combining of a pair of galvo mirrors and a sample stage for changing the resolution and FOV with fixed optical system and objective. The system can be operated not only in low resolution and large FOV, but also in high resolution and small FOV applications, without compromising the performances of the optical elements in the system. By implementing dedicated system control protocol and image reconstruction algorithm, fluorescence images in different FOV and resolution can be obtained according to user’s definition. The performance of the system is demonstrated with two-photon excitation fluorescence imaging of a fluorescence resolution test target, prepared plant stem slide and fluorescence microspheres suspension. Preliminary results indicate that the system has high spatial resolution and holds potential for providing high image acquisition rate. This FOV zoom scanning protocol can be used in our simultaneous time- and spectrum-resolved MMM system, which can provide spectral and lifetime information simultaneously in fluorescence microscopy for biomedical imaging.

© 2007 SPIE

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