OSA's Digital Library

Optics Letters

Optics Letters


  • Vol. 23, Iss. 9 — May. 1, 1998
  • pp: 655–657

Multifocal multiphoton microscopy

Jörg Bewersdorf, Rainer Pick, and Stefan W. Hell  »View Author Affiliations

Optics Letters, Vol. 23, Issue 9, pp. 655-657 (1998)

View Full Text Article

Acrobat PDF (345 KB)

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



We present a real-time, direct-view multiphoton excitation fluorescence microscope that provides three-dimensional imaging at high resolution. Using a rotating microlens disk, we split the near-infrared light of a mode-locked titanium–sapphire laser into an array of beams that are transformed into an array of high-aperture foci at the object. We typically scan at 225 frames per second and image the fluorescence with a camera that reads out the images at video rate. For 1.4 aperture oil and 1.2 water immersion lenses at 780-nm excitation we obtained axial resolutions of 0.84 and 1.4μm , respectively, which are similar to that of a single-beam two-photon microscope. Compared with the latter setup, our system represents a 40–100-fold increase in efficiency, or imaging speed. Moreover, it permits the observation with the eye of high-resolution two-photon images of (live) samples.

© 1998 Optical Society of America

OCIS Codes
(100.6890) Image processing : Three-dimensional image processing
(180.2520) Microscopy : Fluorescence microscopy
(190.4180) Nonlinear optics : Multiphoton processes

Jörg Bewersdorf, Rainer Pick, and Stefan W. Hell, "Multifocal multiphoton microscopy," Opt. Lett. 23, 655-657 (1998)

Sort:  Author  |  Year  |  Journal  |  Reset


  1. W. Denk, J. H. Strickler, and W. W. Webb, Science 248, 73 (1990).
  2. K. König, P. C. T. So, W. W. Mantulin, B. J. Tromberg, and E. Gratton, J. Microsc. 183, 197 (1996).
  3. G. J. Brakenhoff, J. Squier, T. Norris, A. C. Bliton, M. H. Wade, and B. Athey, J. Microsc. 181, 253 (1996).
  4. H. J. Tiziani, R. Achi, R. N. Krämer, and L. Wiegers, Appl. Opt. 35, 120 (1996).
  5. A. Ichihara, T. Tanaami, K. Isozaki, Y. Sugiyama, Y. Kosugi, K. Mikuriya, M. Abe, and I. Uemura, Bioimages 4, 52 (1996).
  6. M. Schrader, U. G. Hofmann, and S. W. Hell, “Ultrathin fluorescent layers for monitoring the axial resolution in confocal and two-photon fluorescence microscopy,” J. Microsc. (to be published).
  7. S. W. Hell, P. E. Hänninen, A. Kuusisto, M. Schrader, and E. Soini, Opt. Commun. 117, 20 (1995).
  8. S. W. Hell, K. Bahlmann, M. Schrader, A. Soini, H. Malak, I. Gryczynski, and J. R. Lakowicz, J. Biomed. Opt. 1, 71 (1996).
  9. D. L. Wokosin, V. E. Centonze, S. Crittenden, and J. White, Bioimaging 4, 208 (1996).

Cited By

Alert me when this paper is cited

OSA is able to provide readers links to articles that cite this paper by participating in CrossRef's Cited-By Linking service. CrossRef includes content from more than 3000 publishers and societies. In addition to listing OSA journal articles that cite this paper, citing articles from other participating publishers will also be listed.

Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited