Simultaneous multiphoton imaging of calcium dynamics and sarcomere contraction on primary cardiomyocytes isolated from transgenic zebrafish
1Department of Applied Chemistry, National Chiao Tung University, Hsinchu, Taiwan
* Presenter:YU KAI CHAO,
Intracelluar calcium ion plays a critical role in regulating the contraction of cardiomyocytes and hence cardiac rhythm.To advance our understanding of its pathophysiological relevance to heart diseases, studying the dynamics between calcium flow and the contraction at the level of single cell is important.By applying multiphoton line-scan imaging on cardiomyocytes isolated from the heart of transgenic line zebrafish Tg(myl7:GCaMP), we reveal delicate dynamics of the intracellular calcium ion fluctuation during cardiomyocyte contraction with no need to introduce exogenous fluorescent probes. Facilitated with MATLAB code, we determined the time-varying sarcomere length, defined as the distance between two adjacent Z-disks, from the second-harmonic generation images of sarcomeric thick filament. For drug response, epinephrine was added to enhance the contractility also the calcium concentration of cardiomyocytes. Overall, we report a method to monitor the dynamic of intracellular calcium ion and sarcomere contraction of zebrafish cardiomyocyte isolated from adult transgenic zebrafish under high spatial-temporal resolution with multiphoton microscopy. We envisage such information could be very useful on fundamental studies or predicting the pharmaceutical effects of new drug during therapeutic intervention.

Keywords: Multiphoton microscopy, Zebrafish, Primary cultured cardiomyocyte