Continuous wave two-photon scanning near-field optical microscopy

被引:20
作者
Kirsch, AK [1 ]
Subramaniam, V [1 ]
Striker, G [1 ]
Schnetter, C [1 ]
Arndt-Jovin, DJ [1 ]
Jovin, TM [1 ]
机构
[1] Max Planck Inst Biophys Chem, Dept Mol Biol, D-37077 Gottingen, Germany
关键词
D O I
10.1016/S0006-3495(98)74070-6
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
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.
引用
收藏
页码:1513 / 1521
页数:9
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