Abstract
Optical mapping (OM) of electrical activity using voltage-sensitive fluorescent dyes is a powerful tool for the investigation of embryonic cardiac electrophysiology. However, because conventional OM integrates the signal in depth and projects it to a two-dimensional plane, information acquired is incomplete and dependent upon the orientation of the sample. This complicates interpretation of data, especially when comparing one heart to another. To overcome this limitation, we present volumetric OM using light-sheet microscopy, which enables high-speed capture of optically sectioned slices. Voltage-sensitive fluorescence images from multiple planes across entire early embryonic quail hearts were acquired, and complete, orientation-independent, four-dimensional maps of transmembrane potential are demonstrated. Volumetric OM data were collected while using optical pacing to control the heart rate, paving the way for physiological measurements and precise manipulation of the heartbeat in the future.
Highlights
During heart development, cardiac electrical signals are important for coordinating the heartbeat as well as providing cues for the normal progression of cardiac development [1, 2]
Abnormal conduction during heart development can lead to congenital heart defects (CHDs) and abnormalities, including defects of the adult cardiac conduction system, abnormal heart morphology, and abnormal blood flow [1, 3, 4]
The bottom side is near the atrial region of the heart and the top side is in the outflow tract
Summary
Cardiac electrical signals are important for coordinating the heartbeat as well as providing cues for the normal progression of cardiac development [1, 2]. With acquired action potential traces, parameters such as activation time, action potential duration and conduction velocity can be computed. With these features, OM has enabled many contributions to the understanding of both embryonic and adult cardiac electrophysiology [7,8,9]. Encoded voltage indicators (GEVIs) are alternatives to voltage-sensitive dyes to target specific cell types and reduce phototoxicity [10]. They are still under development and their applications are limited [10]
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