Multimodal second harmonic generation and two photon fluorescence imaging of microdomain calcium contraction coupling in single cardiomyocytes

James W Chan, Samir Awasthi, Leighton T Izu, Ziliang Mao, Zhong Jian, Trevor Landas, Aaron Lerner, Rafael Shimkunas, Rahwa Woldeyesus, Julie B C Bossuyt, Brittani Wood, Yi-Je Chen, Dennis L Matthews, Deborah Lieu, Nipavan Chiamvimonvat, Kit Lam, Ye Chen-Izu

Research output: Chapter in Book/Report/Conference proceedingConference contribution

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.

Original languageEnglish (US)
Title of host publicationSPIE BioPhotonics Australasia
PublisherSPIE
Volume10013
ISBN (Electronic)9781510604346
DOIs
StatePublished - 2016
EventSPIE BioPhotonics Australasia - Adelaide, Australia
Duration: Oct 17 2016Oct 19 2016

Other

OtherSPIE BioPhotonics Australasia
CountryAustralia
CityAdelaide
Period10/17/1610/19/16

Keywords

  • calcium signaling
  • cardiomyocytes
  • multimodal imaging
  • multiphoton
  • sarcomeres
  • second harmonic generation
  • two photon fluorescence

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Condensed Matter Physics
  • Computer Science Applications
  • Applied Mathematics
  • Electrical and Electronic Engineering

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