TY - JOUR TI - An adaptation of astronomical image processing enables characterization and functional 3D mapping of individual sites of excitation-contraction coupling in rat cardiac muscle AU - Tian, Qinghai AU - Kaestner, Lars AU - Schröder, Laura AU - Guo, Jia AU - Lipp, Peter A2 - Lewis, Richard S VL - 6 PY - 2017 DA - 2017/11/14 SP - e30425 C1 - eLife 2017;6:e30425 DO - 10.7554/eLife.30425 UR - https://doi.org/10.7554/eLife.30425 AB - In beating cardiomyocytes, synchronized localized Ca2+ transients from thousands of active excitation-contraction coupling sites (ECC couplons) comprising plasma and sarcoplasmic reticulum membrane calcium channels are important determinants of the heart's performance. Nevertheless, our knowledge about the properties of ECC couplons is limited by the lack of appropriate experimental and analysis strategies. We designed CaCLEAN to untangle the fundamental characteristics of ECC couplons by combining the astronomer's CLEAN algorithm with known properties of calcium diffusion. CaCLEAN empowers the investigation of fundamental properties of ECC couplons in beating cardiomyocytes without pharmacological interventions. Upon examining individual ECC couplons at the nanoscopic level, we reveal their roles in the negative amplitude-frequency relationship and in β-adrenergic stimulation, including decreasing and increasing firing reliability, respectively. CaCLEAN combined with 3D confocal imaging of beating cardiomyocytes provides a functional 3D map of active ECC couplons (on average, 17,000 per myocyte). CaCLEAN will further enlighten the ECC-couplon-remodelling processes that underlie cardiac diseases. KW - calcium signalling KW - cellular cardiac physiology KW - excitation contraction coupling KW - realtime confocal microscopy JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -