Spontaneous Calcium Transients in iPSC-derived Cardiac Myocytes

Dr. Masayuki Yazawa, a postdoctoral fellow in the Ricardo Dolmetsch lab, took advantage of the environmental control system and fast line scanning capability of the LSM510 Meta confocal to image these spontaneous calcium transients with Fluo4 AM. These data are from human cardiomyocytes generated from induced pluripotent stem cells (iPSC), derived from normal human dermal fibroblasts.

Dr. Yazawa's calcium imaging data from iPSC-derived cardiac myocytes was part of a recent Nature publication, Using induced pluripotent stem cells to investigate cardiac phenotypes in Timothy syndrome. The work was also featured in a Stanford Report news article.

spontaneous calcium transients in iPSC-derived human cardiac myocytes: image data from Dr. Masayuki Yazawa
Image data collected on NMS's LSM510 Meta confocal by Dr. Masayuki Yazawa.

Panel A shows a single cultured cardiac myoctye, loaded with Fluo4 AM. The overlay in red shows the single line that was scanned to monitor changes in calcium levels over time. A single line scan trace (blue) is also shown in the overlay. The length of the scan line is 70 μm. Panel B shows a series of 3700 sequential line scans with time increasing to the right. Panel C shows the transient influxes of calcium as the cultured myocyte spontaneously beats. Each point plotted is the average background-subtracted Fluo4 fluorescence intensity of a single line scan.

As we do for every customer, NMS provided one-on-one training on the scope, and follow-up assistance when needed as the experiments proceeded. Dr. Yazawa writes, "I am now analyzing the data of the calcium imaging from the confocal microscope using Fiji, and I believe that I don't need any more experiments to prepare figures to submit a paper. Thank you so much for your kind help with my experiments. Without your help, my project would have been delayed."