2‐photon excitation fluorescence microscopy enables deeper high‐resolution imaging of voltage and Ca2+ in intact mice,rat, and rabbit hearts |
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| Authors: | Iffath A. Ghouri Allen Kelly Francis L. Burton Godfrey L. Smith Ole Johan Kemi |
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| Affiliation: | 1. Institute of Cardiovascular and Medical Sciences;2. College of Medical, Veterinary, and Life Sciences, University of Glasgow, Glasgow, UK |
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| Abstract: | We describe a novel two‐photon (2P) laser scanning microscopy (2PLSM) protocol that provides ratiometric transmural measurements of membrane voltage (Vm) via Di‐4‐ANEPPS in intact mouse, rat and rabbit hearts with subcellular resolution. The same cells were then imaged with Fura‐2/AM for intracellular Ca2+ recordings. Action potentials (APs) were accurately characterized by 2PLSM vs. microelectrodes, albeit fast events (<1 ms) were sub‐optimally acquired by 2PLSM due to limited sampling frequencies (2.6 kHz). The slower Ca2+ transient (CaT) time course (>1ms) could be accurately described by 2PLSM. In conclusion, Vm ‐ and Ca2+‐sensitive dyes can be 2P excited within the cardiac muscle wall to provide AP and Ca2+ signals to ～400 µm. (© 2013 WILEY‐VCH Verlag GmbH &Co. KGaA, Weinheim) |
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| Keywords: | action potential |
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