We have developed a high-speed two-photon microscope with submicrometer resolution in real time. The imaging speed improvement of this system is obtained by the use of a high-speed polygonal mirror scanner. The maximum achievable scanning rate is 40 μs/line, which is approximately 100 times faster than conventional scanning microscopes. High-resolution fluorescence images were recorded in real time by an intensified CCD camera. Using this instrument, we have resolved cellular architecture in three dimensions and have monitored the movements of protozoas. More important, photodamage to biological specimens during video-rate imaging can be minimized with two-photon excitation as compared with other one-photon modalities.
© 1999 Optical Society of America
(110.0180) Imaging systems : Microscopy
(120.3890) Instrumentation, measurement, and metrology : Medical optics instrumentation
(170.0110) Medical optics and biotechnology : Imaging systems
(170.5810) Medical optics and biotechnology : Scanning microscopy
(180.2520) Microscopy : Fluorescence microscopy
Ki Hean Kim, Christof Buehler, and Peter T. C. So, "High-Speed, Two-Photon Scanning Microscope," Appl. Opt. 38, 6004-6009 (1999)