title: Pulse splitter-based nonlinear microscopy for live-cardiomyocyte imaging.
abstract: Second harmonic generation (SHG) microscopy is a new imaging technique used in sarcomeric-addition studies. However, during the early stage of cell culture in which sarcomeric additions occur, the neonatal cardiomyocytes that we have been working with are very sensitive to photodamage, the resulting high rate of cell death prevents systematic study of sarcomeric addition using a conventional SHG system. To address this challenge, we introduced use of the pulse-splitter system developed by Na Ji et al. in our two photon excitation fluorescence (TPEF) and SHG hybrid microscope. The system dramatically reduced photodamage to neonatal cardiomyocytes in early stages of culture, greatly increasing cell viability. Thus continuous imaging of live cardiomyocytes was achieved with a stronger laser and for a longer period than has been reported in the literature. The pulse splitter-based TPEF-SHG microscope constructed in this study was demonstrated to be an ideal imaging system for sarcomeric addition-related investigations of neonatal cardiomyocytes in early stages of culture.
authors.affiliation: Department of Bioengineering, COMSET, Clemson University, Clemson, SC 29631, USA.
authors.forename: Zhonghai
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Department of Bioengineering, COMSET, Clemson University, Clemson, SC 29631, USA.
authors.forename: Wan
authors.initials: W
authors.lastname: Qin
authors.affiliation: Institute of Optoelectronics, Shenzhen University, Shenzhen, Guangdong 518060, PR China.
authors.forename: Yonghong
authors.initials: Y
authors.lastname: Shao
authors.affiliation: Department of Bioengineering, COMSET, Clemson University, Clemson, SC 29631, USA.
authors.forename: Siyu
authors.initials: S
authors.lastname: Ma
authors.affiliation: Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, SC 29425, USA.
authors.forename: Thomas K
authors.initials: TK
authors.lastname: Borg
authors.affiliation: Department of Bioengineering, COMSET, Clemson University, Clemson, SC 29631, USA.
authors.forename: Bruce Z
authors.initials: BZ
authors.lastname: Gao
keywords: SHG; TPEF; cardiomyocyte; hypertrophy; live imaging; pulse-splitter