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Proceedings Paper

Second- and third-harmonic generation and multiphoton excitation fluorescence microscopy for simultaneous imaging of cardiomyocytes
Author(s): Virginijus Barzda; Catherine Greenhalgh; Juerg Aus der Au; Jeffrey A. Squier; Steven Elmore; Johannes H.G.M. van Beek
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Paper 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.

Paper Details

Date Published: 1 June 2004
PDF: 8 pages
Proc. SPIE 5340, Commercial and Biomedical Applications of Ultrafast Lasers IV, (1 June 2004); doi: 10.1117/12.529287
Show Author Affiliations
Virginijus Barzda, Univ. of Toronto (Canada)
Catherine Greenhalgh, Univ. of Toronto (Canada)
Juerg Aus der Au, Colorado School of Mines (United States)
Jeffrey A. Squier, Colorado School of Mines (United States)
Steven Elmore, Vrije Univ. Brussel (Belgium)
Johannes H.G.M. van Beek, Vrije Univ. Brussel (Belgium)

Published in SPIE Proceedings Vol. 5340:
Commercial and Biomedical Applications of Ultrafast Lasers IV
Joseph Neev; Christopher B. Schaffer; Andreas Ostendorf, Editor(s)

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