24 November 2016 Multimodal second harmonic generation and two photon fluorescence imaging of microdomain calcium contraction coupling in single cardiomyocytes
Author Affiliations +
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.
© (2016) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
James Chan, James Chan, Samir Awasthi, Samir Awasthi, Leighton Izu, Leighton Izu, Ziliang Mao, Ziliang Mao, Zhong Jian, Zhong Jian, Trevor Landas, Trevor Landas, Aaron Lerner, Aaron Lerner, Rafael Shimkunas, Rafael Shimkunas, Rahwa Woldeyesus, Rahwa Woldeyesus, Julie Bossuyt, Julie Bossuyt, Brittani Wood, Brittani Wood, Yi-Je Chen, Yi-Je Chen, Dennis Matthews, Dennis Matthews, Deborah Lieu, Deborah Lieu, Nipavan Chiamvimonvat, Nipavan Chiamvimonvat, Kit Lam, Kit Lam, Ye Chen-Izu, 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); doi: 10.1117/12.2242805; https://doi.org/10.1117/12.2242805
PROCEEDINGS
7 PAGES


SHARE
Back to Top