Second- and third-harmonic generation and multiphoton excitation fluorescence microscopy for simultaneous imaging of cardiomyocytes

Virginijus Barzda; Catherine Greenhalgh; Juerg Aus der Au; Jeffrey A. Squier; Steven Elmore; Johannes H.G.M. van Beek

doi:10.1117/12.529287

1 June 2004 Second- and third-harmonic generation and multiphoton excitation fluorescence microscopy for simultaneous imaging of cardiomyocytes

Virginijus Barzda, Catherine Greenhalgh, Juerg Aus der Au, Jeffrey A. Squier, Steven Elmore, Johannes H.G.M. van Beek

Author Affiliations +

Proceedings Volume 5340, Commercial and Biomedical Applications of Ultrafast Lasers IV; (2004) https://doi.org/10.1117/12.529287
Event: Lasers and Applications in Science and Engineering, 2004, San Jose, Ca, United States

Abstract

Simultaneous detection of second harmonic generation (SHG), third harmonic generation (THG) and multiphoton excitation fluorescence with ultrafast laser pulses from a Nd:Glass laser was used to image isolated adult rat cardiomyocytes. The simultaneous detection enabled visualization of different organelles of cardiomyocytes, based on the different contrast mechanisms. It was found that SHG signal depicted characteristic patterns of sarcomeres in a myofilament lattice. The regular pattern of the THG signal, which was anticorrelated with the SHG signal, suggested that the third harmonic is generated within mitochondria. By labeling the cardiomyocytes with the mitochondrial dye tetramethylrhodamine methyl ester (TMRM), comparisons could be made between the TMRM fluorescence, THG, and SHG images. The TMRM fluorescence had significant correlation with THG signal confirming that part of the THG signal originates from mitochondria.

Citation Download Citation

Virginijus Barzda, Catherine Greenhalgh, Juerg Aus der Au, Jeffrey A. Squier, Steven Elmore, and Johannes H.G.M. van Beek "Second- and third-harmonic generation and multiphoton excitation fluorescence microscopy for simultaneous imaging of cardiomyocytes", Proc. SPIE 5340, Commercial and Biomedical Applications of Ultrafast Lasers IV, (1 June 2004); https://doi.org/10.1117/12.529287

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