Functional imaging of primary neonatal mouse cardiomyocytes using extended depth-of-field microscopy

Abstract

The recent development of binary phase filters (BPFs) has achieved the ability to extend the depth-of-field (DoF) of optical systems while maintaining high resolution, demonstrating the anticipated impact of BPF-induced extended depth-of-field (eDoF) microscopy in the biological and medical fields. To suggest the novel application of BPF-induced eDoF to functional imaging, this study applied a BPF-induced eDoF imaging system to observe primary neonatal mouse cardiomyocytes. Specifically, calcium imaging with the fluorescent dye Fluo-3 AM was utilized to monitor cardiomyocyte calcium transients, which serve as key indicators of cardiomyocyte function and viability. Findings show that Fluo-3 AM calcium imaging with BPF-induced eDoF microscopy enables precise visualization and analysis of calcium dynamics in primary neonatal mouse cardiomyocytes, although with needed areas of improvement. This study’s application of a BPF-induced eDoF technique on the functional imaging of primary cardiomyocyte cultures paves way for further improved imaging-based analysis while also underscoring the importance of utilizing such advanced tools.