Live imaging of contracting muscles with wide-field second harmonic generation microscopy using a high power laser

Haitao Zhao; Richard Cisek; Abiramy Karunendiran; Danielle Tokarz; Bryan A Stewart; Virginijus Barzda

doi:10.1364/BOE.10.005130

Live imaging of contracting muscles with wide-field second harmonic generation microscopy using a high power laser

Biomed Opt Express. 2019 Sep 13;10(10):5130-5135. doi: 10.1364/BOE.10.005130. eCollection 2019 Oct 1.

Authors

Haitao Zhao^{1

2

3

4}, Richard Cisek^{5

4}, Abiramy Karunendiran^{6

7}, Danielle Tokarz⁵, Bryan A Stewart^{6

7}, Virginijus Barzda^{2

3}

Affiliations

¹ WDI Device Inc., 135 West Beaver Creek Road Unit 2, Richmond Hill, ON, L4B 1L2, Canada.
² Department of Chemical and Physical Sciences, University of Toronto Mississauga, 3359 Mississauga Road, Mississauga, ON L5L 1C6, Canada.
³ Department of Physics, University of Toronto, 60 St. George Street, Toronto, ON M5S 1A7, Canada.
⁴ Co-first authors with equal contribution.
⁵ Department of Chemistry, Saint Mary's University, 923 Robie Street, Halifax, NS B3H 3C3, Canada.
⁶ Department of Biology, University of Toronto Mississauga, 3359 Mississauga Road, Mississauga, ON L5L 1C6, Canada.
⁷ Department of Cell and Systems Biology, University of Toronto, 25 Harbord Street, Toronto, ON, M5S 3G5, Canada.

Abstract

Wide-field second harmonic generation (SHG) microscopy was developed using a high-power (> 4 W) and high-repetition-rate (MHz range) laser oscillator to achieve fast SHG imaging over a large area (400 µm × 400 µm). The microscope was used for high spatial resolution imaging of contracting muscles in live Drosophila melanogaster larvae. Anisotropic and isotropic bands of striated muscle were distinguished, allowing accurate determination of sarcomere length and SHG intensity from individual sarcomeres. Therefore, wide-field SHG microscopy has applications in basic contractility research and studying arrhythmias, muscular dystrophies and pharmaceutical effects on the muscle contraction dynamics of sarcomeres.