Optogenetic stimulation and simultaneous optical mapping of membrane potential and calcium transients in human engineered cardiac spheroids.

ABSTRACT: Optogenetic stimulation combined with optical mapping of membrane potential (Vm) and calcium transients (CaT) is a powerful electrophysiological tool. We developed a novel experimental platform in which tissue is stimulated optogenetically while Vm and CaT are imaged simultaneously. The Vm indicator is an organic dye, while the CaT indicator is genetically encoded. We used cardiac spheroids containing cardiomyocytes and fibroblasts differentiated from human induced pluripotent stem cells as model tissue. The spheroids were genetically encoded with an optogenetic actuator, CheRiff, and the calcium indicator jRCaMP1b. The Vm indicator was the organic dye RH237. CheRiff was excited using blue light (450 nm), and both RH237 and jRCaMP1b were excited using a single band of green light (either 5

SUBMITTER: Guragain B 

PROVIDER: S-EPMC11788028 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature