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Applied Optics

Applied Optics


  • Vol. 38, Iss. 28 — Oct. 1, 1999
  • pp: 6004–6009

High-speed, two-photon scanning microscope

Ki Hean Kim, Christof Buehler, and Peter T. C. So  »View Author Affiliations

Applied Optics, Vol. 38, Issue 28, pp. 6004-6009 (1999)

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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

OCIS Codes
(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

Original Manuscript: November 30, 1998
Revised Manuscript: June 16, 1999
Published: October 1, 1999

Ki Hean Kim, Christof Buehler, and Peter T. C. So, "High-speed, two-photon scanning microscope," Appl. Opt. 38, 6004-6009 (1999)

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