2-photon excitation fluorescence microscopy enables deeper high-resolution imaging of voltage and Ca2+in intact mice, rat, and rabbit hearts

Ghouri, I. A., Kelly, A., Burton, F. L., Smith, G. L. and Kemi, O. J. (2015) 2-photon excitation fluorescence microscopy enables deeper high-resolution imaging of voltage and Ca2+in intact mice, rat, and rabbit hearts. Journal of Biophotonics, 8(1-2), pp. 112-123. (doi: 10.1002/jbio.201300109)

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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.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Burton, Dr Francis and Smith, Professor Godfrey and Ghouri, Dr Iffath and Kelly, Dr Allen and Kemi, Dr Ole
Authors: Ghouri, I. A., Kelly, A., Burton, F. L., Smith, G. L., and Kemi, O. J.
College/School:College of Medical Veterinary and Life Sciences > Institute of Cardiovascular and Medical Sciences
Journal Name:Journal of Biophotonics
Publisher:WILEY-VCH Verlag GmbH & Co. KGaA
ISSN:1864-063X
ISSN (Online):1864-0648
Copyright Holders:Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA
First Published:First published in Journal of Biophotonics 8(1-2):112-123
Publisher Policy:Reproduced in accordance with the copyright policy of the publisher.

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