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Journal of Biomedical Optics • Open Access

Multispot multiphoton Ca2+ imaging in acute myocardial slices
Author(s): Giulia Borile; Claudio de Mauro; Andrea Urbani; Domenico Alfieri; Francesco S. Pavone; Marco Mongillo

Paper Abstract

Multiphoton microscopy has become essential for dynamic imaging in thick living tissues. High-rate, full-field image acquisition in multiphoton microscopy is achievable by parallelization of the excitation and detection pathways. We developed our approach via a diffractive optical element which splits a pulsed laser into 16 beamlets and exploits a descanned detection system consisting of an array of beamlet-associated photomultiplier tubes. The optical performance of the multiphoton multispot system (MCube) has been characterized in cardiac tissue sections and subsequently used for the first time for fluorescence imaging of cardiomyocyte Ca2+ dynamics in viable acute cardiac slices. Multispot multiphoton microscopy (MMM) has never been used before to monitor Ca2+ dynamics in thick, viable tissue samples. Acute heart slices are a powerful close-to-in vivo model of Ca2+ imaging allowing the simultaneous observation of several cells in their own tissue environment, exploiting the multiphoton excitation ability to penetrate scattering tissues. Moreover, we show that the concurrent high spatial and temporal resolutions afforded by the parallel scanning in MMM can be exploited to simultaneously assess subcellular Ca2+ dynamics in different cells in the tissue. We recorded local Ca2+ release events including macrosparks, travelling waves, and rotors.

Paper Details

Date Published: 17 December 2014
PDF: 8 pages
J. Biomed. Opt. 20(5) 051016 doi: 10.1117/1.JBO.20.5.051016
Published in: Journal of Biomedical Optics Volume 20, Issue 5
Show Author Affiliations
Giulia Borile, Univ. degli Studi di Padova (Italy)
Venetian Institute of Molecular Medicine (Italy)
Claudio de Mauro, Light4Tech Firenze S.r.l. (Italy)
Andrea Urbani, Univ. of Padova (Italy)
Venetian Institute of Molecular Medicine (Italy)
Domenico Alfieri, Light4Tech Firenze S.r.l. (Italy)
Francesco S. Pavone, Univ. of Florence (Italy)
Marco Mongillo, Univ. degli Studi di Padova (Italy)
Venetian Institute of Molecular Medicine (Italy)

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