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Proceedings Paper

Study of the sarcomeric addition process in a tissue-like cell construct under mechanical overload via TPEF-SHG imaging system
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Paper Abstract

Using our lab built two-photon excitation fluorescence (TPEF) and second harmonic generation (SHG) hybrid confocal imaging system, we observed, for the first time, the dynamic sarcomeric addition process in a rat cardiomyocyte cell culture system; this culture system expressed in vivo-like myofibril structure and mimicked mechanical overload experienced in a heart muscle tissue. Micro-grooved topographic patterned substrates com- bined with electrical stimulation are used to achieve the in vivo-like myofibril structure. After cardiomyocytes aligned, longitudinal and transverse mechanical stretch was applied to cardiomyocytes in parallel or perpendicu- lar, respectively, to the direction of alignment via stretching the substrates to mimic mechanical overload. Z discs, in which alpha-actinin expressed, have been proposed to involve in the process. TPEF detected alpha-actinin that labeled with enhanced yellow florescent protein via plasmid transfection. SHG is intrinsic to noncentrosym- metric structures, thus was used to detect myosin, a polar molecule expressed in myofibril. Pulse splitter system and synchronized recording system was introduced on TPEF-SHG imaging system to reduce the photodamage during live cell imaging. In our study, TPEF-SHG imaging system was used to study the dynamic process of sarcomeric addition in in vivo-like culture model under mechanical overload. This microscopic technique is ideal for tracking sarcomeric components to successively assemble onto pre-exist myofibrils and for revealing the role of Z discs played in sarcomeric addition. Transition of Z discs from continuous to broadened striation and from broadened to uniform striation under stretch has been observed. We concluded that continuous Z discs is the place of new sarcomeric addition.

Paper Details

Date Published: 14 February 2020
PDF: 10 pages
Proc. SPIE 11244, Multiphoton Microscopy in the Biomedical Sciences XX, 112442E (14 February 2020); doi: 10.1117/12.2546280
Show Author Affiliations
Ailin Wei, Clemson Univ. (United States)
Zhonghai Wang, Clemson Univ. (United States)
Zongming Yang, Clemson Univ. (United States)
Shenghao Tan, Clemson Univ. (United States)
Tong Ye, Clemson Univ. (United States)
Yonghong Shao, Shenzhen Univ. (China)
Thomas K. Borg, Medical Univ. of South Carolina (United States)
Bruce Z. Gao, Clemson Univ. (United States)

Published in SPIE Proceedings Vol. 11244:
Multiphoton Microscopy in the Biomedical Sciences XX
Ammasi Periasamy; Peter T. C. So; Karsten König, Editor(s)

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