Simultaneous multichannel nonlinear imaging: combined two-photon excited fluorescence and second-harmonic generation microscopy

Regis Gauderon; Philip B. Lukins; Colin J. R. Sheppard

doi:10.1117/12.390533

4 July 2000 Simultaneous multichannel nonlinear imaging: combined two-photon excited fluorescence and second-harmonic generation microscopy

Regis Gauderon, Philip B. Lukins, Colin J. R. Sheppard

Author Affiliations +

Proceedings Volume 4082, Optical Sensing, Imaging, and Manipulation for Biological and Biomedical Applications; (2000) https://doi.org/10.1117/12.390533
Event: Photonics Taiwan, 2000, Taipei, Taiwan

Abstract

Simultaneous two-photo excited fluorescence (TPF) and second- harmonic generation (SHG) imaging is demonstrated using a single femtosecond laser and a scanning microscope. This composite nonlinear microscopic technique was applied to imaging DNA and chromosomes, and it was shown that the two different interaction mechanisms provide complementary information on the structure and nonlinear properties of these biological materials beyond that achievable using either TPF or SHG imaging alone. The use of separate modes of detection, in reflection and transmission respectively, and the simultaneous nature of the acquisition of the two images allows pure TPF and SHG images in precise registration to be obtained.

Citation Download Citation

Regis Gauderon, Philip B. Lukins, and Colin J. R. Sheppard "Simultaneous multichannel nonlinear imaging: combined two-photon excited fluorescence and second-harmonic generation microscopy", Proc. SPIE 4082, Optical Sensing, Imaging, and Manipulation for Biological and Biomedical Applications, (4 July 2000); https://doi.org/10.1117/12.390533

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