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Optics Letters

Optics Letters

| RAPID, SHORT PUBLICATIONS ON THE LATEST IN OPTICAL DISCOVERIES

  • Vol. 23, Iss. 15 — Aug. 1, 1998
  • pp: 1238–1240

Two-photon near- and far-field fluorescence microscopy with continuous-wave excitation

Stefan W. Hell, Martin Booth, Stefan Wilms, Christoph M. Schnetter, Achim K. Kirsch, Donna J. Arndt-Jovin, and Thomas M. Jovin  »View Author Affiliations


Optics Letters, Vol. 23, Issue 15, pp. 1238-1240 (1998)
http://dx.doi.org/10.1364/OL.23.001238


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Abstract

We report on scanning far- and near-field two-photon microscopy of cell nuclei stained with DAPI and bisbenzimidazole Hoechst 33342 (BBI-342) with the 647-nm laser line of a cw ArKr mixed-gas laser. Two-photon-excited fluorescence images are obtained for 50–200 mW of average power at the sample. A nearly quadratic dependence of fluorescence intensity on laser power confirmed the two-photon effect. The nonlinearity was further supported by evidence of three-dimensional sectioning in a scanning far-field microscope. We find that the cw two-photon irradiation sufficient for imaging within typically 5 s does not significantly impair cell cycling of BBI-342-labeled live cells. Finally, high-resolution imaging in scanning near-field microscopy with good contrast is demonstrated.

© 1998 Optical Society of America

OCIS Codes
(140.3460) Lasers and laser optics : Lasers
(170.0180) Medical optics and biotechnology : Microscopy
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(170.5280) Medical optics and biotechnology : Photon migration

Citation
Stefan W. Hell, Martin Booth, Stefan Wilms, Christoph M. Schnetter, Achim K. Kirsch, Donna J. Arndt-Jovin, and Thomas M. Jovin, "Two-photon near- and far-field fluorescence microscopy with continuous-wave excitation," Opt. Lett. 23, 1238-1240 (1998)
http://www.opticsinfobase.org/ol/abstract.cfm?URI=ol-23-15-1238

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