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24 November 2016 Multimodal second harmonic generation and two photon fluorescence imaging of microdomain calcium contraction coupling in single cardiomyocytes
James Chan, Samir Awasthi, Leighton Izu, Ziliang Mao, Zhong Jian, Trevor Landas, Aaron Lerner, Rafael Shimkunas, Rahwa Woldeyesus, Julie Bossuyt, Brittani Wood, Yi-Je Chen, Dennis Matthews, Deborah Lieu, Nipavan Chiamvimonvat M.D., Kit Lam, Ye Chen-Izu
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Proceedings Volume 10013, SPIE BioPhotonics Australasia; 1001304 (2016) https://doi.org/10.1117/12.2242805
Event: SPIE BioPhotonics Australasia, 2016, Adelaide, Australia
Abstract
The objective of this study was to develop a method for simultaneously measuring the calcium and contraction dynamics of single, live cardiomyocytes at high spatial resolutions. Such measurements are important to investigate local calcium release and the mechanical response at the sarcomere level (i.e. the basic unit of contraction), which have important implications in cardiac dysfunction and arrhythmias in conditions such as hypertension, atrial fibrillation, and myocardial infarction. Here, we describe a multimodal second harmonic generation (SHG) and two photon fluorescence (2PF) microscopy technique that is used to simultaneously measure subsarcomere calcium and contraction events at high spatial and temporal resolutions. The method takes advantage of the label-free nature of SHG for imaging the sarcomeres and the high spatial colocalization of the SHG signal and the fluorescence signal excited from calcium indicators. This microscope was used to measure calcium sparks and waves and associated contractions in subcellular microdomains, leading to the generation of subcellular strain. We anticipate this new imaging tool will play an important role in studying mechanical stress-induced heart disease.
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James Chan, Samir Awasthi, Leighton Izu, Ziliang Mao, Zhong Jian, Trevor Landas, Aaron Lerner, Rafael Shimkunas, Rahwa Woldeyesus, Julie Bossuyt, Brittani Wood, Yi-Je Chen, Dennis Matthews, Deborah Lieu, Nipavan Chiamvimonvat M.D., Kit Lam, and Ye Chen-Izu "Multimodal second harmonic generation and two photon fluorescence imaging of microdomain calcium contraction coupling in single cardiomyocytes", Proc. SPIE 10013, SPIE BioPhotonics Australasia, 1001304 (24 November 2016); https://doi.org/10.1117/12.2242805
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KEYWORDS
Calcium
Second-harmonic generation
Luminescence
Heart
Harmonic generation
Microscopes
Two photon imaging
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