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  Continuous wave two-photon scanning near-field optical microscopy

Kirsch, A. K., Subramaniam, V., Striker, G., Schnetter, C., Arndt-Jovin, D. J., & Jovin, T. M. (1998). Continuous wave two-photon scanning near-field optical microscopy. Biophysical Journal, 75, 1513-1521.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0012-FD2B-F Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-EC05-9
Genre: Journal Article

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 Creators:
Kirsch, A. K.1, Author              
Subramaniam, V.1, Author              
Striker, G.1, Author              
Schnetter, C.1, Author              
Arndt-Jovin, D. J.1, Author              
Jovin, T. M.1, Author              
Affiliations:
1Department of Molecular Biology, MPI for biophysical chemistry, Max Planck Society, ou_578628              

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Free keywords: Animal; Biophysics; Cell Nucleus/ul [Ultrastructure]; Chromosomes/ul [Ultrastructure]; Drosophila melanogaster; Fluorescent Dyes; Indoles; Mice; Microscopy, Fluorescence/is [Instrumentation]; *Microscopy, Fluorescence/mt [Methods]; Photons; Support, Non-U.S. Gov't; 3T3 Cells; 2-photon fluorescence microscopy; Living cells; Laser; Protein; Dna; Spectroscopy; Liquids; Dapi
 Abstract: We have implemented continuous-wave two-photon excitation of near-UV absorbing fluorophores in a scanning near-field optical microscope (SNOM). The 647-nm emission of an Ar-Kr mixed gas laser was used to excite the UV-absorbing DNA dyes DAPI, the bisbenzimidazole Hoechst 33342, and ethidium bromide in a shared aperture SNOM with uncoated fiber tips. Polytene chromosomes of Drosophila melanogaster and the nuclei of 3T3 Balb/c cells labeled with these dyes were readily imaged. The fluorescence intensity showed the expected nonlinear (second order) dependence on the excitation power in the range of 8-180 mW. We measured the fluorescence intensity as a function of the tip-sample displacement in the direction normal to the sample surface in the single- and two-photon excitation modes (SPE, TPE). The fluorescence intensity decayed faster in TPE than in SPE.

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Language(s): eng - English
 Dates: 2005-06-241998
 Publication Status: Published in print
 Pages: -
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 Table of Contents: -
 Rev. Method: Peer
 Identifiers: eDoc: 226993
Other: 634
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Title: Biophysical Journal
  Alternative Title : Biophys. J.
Source Genre: Journal
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Pages: - Volume / Issue: 75 Sequence Number: - Start / End Page: 1513 - 1521 Identifier: -