CytoCypher BV, Wageningen, The Netherlands.
Leiden Institute of Advanced Computer Science, University of Leiden, Leiden, The Netherlands.
PLoS One. 2021 Sep 1;16(9):e0256713. doi: 10.1371/journal.pone.0256713. eCollection 2021.
Simultaneous calcium and contractility measurements on isolated adult cardiomyocytes have been the gold standard for the last decades to study cardiac (patho)physiology. However, the throughput of this system is low which limits the number of compounds that can be tested per animal. We developed instrumentation and software that can automatically find adult cardiomyocytes. Cells are detected based on the cell boundary using a Sobel-filter to find the edge information in the field of view. Separately, we detected motion by calculating the variance of intensity for each pixel in the frame through time. Additionally, it detects the best region for calcium and contractility measurements. A sensitivity of 0.66 ± 0.08 and a precision of 0.82 ± 0.03 was reached using our cell finding algorithm. The percentage of cells that were found and had good contractility measurements was 90 ± 10%. In addition, the average time between 2 cardiomyocyte calcium and contractility measurements decreased from 93.5 ± 80.2 to 15.6 ± 8.0 seconds using our software and microscope. This drastically increases throughput and provides a higher statistical reliability when performing adult cardiomyocyte functional experiments.
几十年来,分离的成年心肌细胞的钙和收缩性的同步测量一直是研究心脏(病理)生理学的金标准。然而,该系统的通量较低,限制了每只动物可以测试的化合物数量。我们开发了可以自动找到成年心肌细胞的仪器和软件。细胞是基于细胞边界使用 Sobel 滤波器检测到的,以找到视野中的边缘信息。另外,通过在帧内随时间计算每个像素的强度方差,来检测运动。此外,它还检测钙和收缩性测量的最佳区域。使用我们的细胞检测算法,达到了 0.66 ± 0.08 的灵敏度和 0.82 ± 0.03 的精度。找到并具有良好收缩性测量的细胞百分比为 90 ± 10%。此外,使用我们的软件和显微镜,两次心肌细胞钙和收缩性测量之间的平均时间从 93.5 ± 80.2 秒减少到 15.6 ± 8.0 秒。这大大提高了通量,并在进行成年心肌细胞功能实验时提供了更高的统计可靠性。