Visualization of nanocontractions in cardiomyocytes by simultaneous detection of second- and third- harmonic generation and multiphoton excitation fluorescence microscopy

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

doi:10.1117/12.591212

24 March 2005 Visualization of nanocontractions in cardiomyocytes by simultaneous detection of second- and third- harmonic generation and multiphoton excitation fluorescence microscopy

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

Author Affiliations +

Proceedings Volume 5701, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XII; (2005) https://doi.org/10.1117/12.591212
Event: SPIE BiOS, 2005, San Jose, CA, United States

Abstract

Dynamic imaging of cardiomyocytes was performed with a simultaneous detection second harmonic generation (SHG), third harmonic generation (THG) and multiphoton excitation fluorescence (MPF) microscope. The fast scanning system of ~12 frames/second synchronized with multichannel detection provided the possibility of imaging three dimensional static and two dimensional dynamic structures of cardiomyocytes. The SHG images highlighted the myofibrils of the cardiomyocytes while THG images revealed the locations of mitochondria. Dynamic data showed that during imaging, chaotic nanocontractions took place inside the cardiomyocytes. The time series of THG images reveled large intensity fluctuations "flickering" in the regions of mitochondria. The flickering in THG correlated with the flickering in MPF. Addition of the uncoupler FCCP inhibited flickering in THG and MPF, and also inhibited nanocontractions. The simultaneous imaging with SHG, THG and MPF proved to be a very powerful microscopy tool for investigation of interactions of different organelles inside a cell.

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Catherine Greenhalgh, Virginijus Barzda, Steven Elmore, Johannes H. G. M. van Beek, Juerg Aus der Au, and Jeffrey Squier "Visualization of nanocontractions in cardiomyocytes by simultaneous detection of second- and third- harmonic generation and multiphoton excitation fluorescence microscopy", Proc. SPIE 5701, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XII, (24 March 2005); https://doi.org/10.1117/12.591212

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