KHz-rate volumetric voltage imaging of the whole zebrafish heart

Sacconi, Leonardo; Silvestri, Ludovico; Rodríguez, E.C.; Armstrong, Gary A. B.; Pavone, Francesco S.; Shrier, Alvin; Bub, Gil

doi:10.1101/2020.07.13.196063

Cited by 4 publications

(4 citation statements)

References 32 publications

(30 reference statements)

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“…The resulting images then correspond to a small z-stack in object space. In the simplest form of MPM, multiple non-polarizing 50:50 beamsplitters (BS) can be used in combination with different camera positions offset from the nominal focal plane (as depicted in Figure 3B) or tube lenses with different focal lengths can be used [22,[26][27][28][29][30][31][32]. To decrease the spatial footprint of this configuration, mirrors can be interspaced (Figure 3C) with the BS [12,22].…”

Section: Multiplane Imagingmentioning

confidence: 99%

Mapping volumes to planes: Camera-based strategies for snapshot volumetric microscopy

Engelhardt

Grußmayer

2022

Front. Phys.

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Optical microscopes allow us to study highly dynamic events from the molecular scale up to the whole animal level. However, conventional three-dimensional microscopy architectures face an inherent tradeoff between spatial resolution, imaging volume, light exposure and time required to record a single frame. Many biological processes, such as calcium signalling in the brain or transient enzymatic events, occur in temporal and spatial dimensions that cannot be captured by the iterative scanning of multiple focal planes. Snapshot volumetric imaging maintains the spatio-temporal context of such processes during image acquisition by mapping axial information to one or multiple cameras. This review introduces major methods of camera-based single frame volumetric imaging: so-called multiplane, multifocus, and light field microscopy. For each method, we discuss, amongst other topics, the theoretical framework; tendency towards optical aberrations; light efficiency; applicable wavelength range; robustness/complexity of hardware and analysis; and compatibility with different imaging modalities, and provide an overview of applications in biological research.

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Section: Multiplane Imagingmentioning

confidence: 99%

Mapping volumes to planes: Camera-based strategies for snapshot volumetric microscopy

Engelhardt

Grußmayer

2022

Front. Phys.

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“…Dual excitation of spectrally separated VSD has also been used successfully to reveal unique information about transmural heterogeneities and endocardial electrophysiology in rat hearts (Walton et al, 2010 ). Looking to the future, with the emergence of low-cost high-speed cameras, volumetric optical mapping in small-size hearts has been shown to be possible with multi-plane multi-camera parallel imaging (Sacconi et al, 2020 ).…”

Section: Multimodal Imaging Of Intracellular Ca 2+ and Membrane Potentialmentioning

confidence: 99%

Novel Optics-Based Approaches for Cardiac Electrophysiology: A Review

et al. 2021

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Optical techniques for recording and manipulating cellular electrophysiology have advanced rapidly in just a few decades. These developments allow for the analysis of cardiac cellular dynamics at multiple scales while largely overcoming the drawbacks associated with the use of electrodes. The recent advent of optogenetics opens up new possibilities for regional and tissue-level electrophysiological control and hold promise for future novel clinical applications. This article, which emerged from the international NOTICE workshop in 20181, reviews the state-of-the-art optical techniques used for cardiac electrophysiological research and the underlying biophysics. The design and performance of optical reporters and optogenetic actuators are reviewed along with limitations of current probes. The physics of light interaction with cardiac tissue is detailed and associated challenges with the use of optical sensors and actuators are presented. Case studies include the use of fluorescence recovery after photobleaching and super-resolution microscopy to explore the micro-structure of cardiac cells and a review of two photon and light sheet technologies applied to cardiac tissue. The emergence of cardiac optogenetics is reviewed and the current work exploring the potential clinical use of optogenetics is also described. Approaches which combine optogenetic manipulation and optical voltage measurement are discussed, in terms of platforms that allow real-time manipulation of whole heart electrophysiology in open and closed-loop systems to study optimal ways to terminate spiral arrhythmias. The design and operation of optics-based approaches that allow high-throughput cardiac electrophysiological assays is presented. Finally, emerging techniques of photo-acoustic imaging and stress sensors are described along with strategies for future development and establishment of these techniques in mainstream electrophysiological research.

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“…They discovered two compounds that rescued wild-type-like APD in breakdance larvae, suggesting novel pathways for restoration of repolarization associated with hERG dysfunction. More recently, light-sheet imaging of a membrane dye in whole 14 dpf zebrafish hearts showed that E-4031 (a hERG blocker) application increases APD and induces the occurrence of frequent EAD formation (∼1 every s) ( Sacconi et al, 2020 ). These studies demonstrate the applicability of zebrafish larvae to model acquired LQTS and to conduct toxicological screening.…”

Section: Zebrafish As a Model Of Acquired Long-qt Syndromementioning

confidence: 99%

Utility of Zebrafish Models of Acquired and Inherited Long QT Syndrome

et al. 2021

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Long-QT Syndrome (LQTS) is a cardiac electrical disorder, distinguished by irregular heart rates and sudden death. Accounting for ∼40% of cases, LQTS Type 2 (LQTS2), is caused by defects in the Kv11.1 (hERG) potassium channel that is critical for cardiac repolarization. Drug block of hERG channels or dysfunctional channel variants can result in acquired or inherited LQTS2, respectively, which are typified by delayed repolarization and predisposition to lethal arrhythmia. As such, there is significant interest in clear identification of drugs and channel variants that produce clinically meaningful perturbation of hERG channel function. While toxicological screening of hERG channels, and phenotypic assessment of inherited channel variants in heterologous systems is now commonplace, affordable, efficient, and insightful whole organ models for acquired and inherited LQTS2 are lacking. Recent work has shown that zebrafish provide a viable in vivo or whole organ model of cardiac electrophysiology. Characterization of cardiac ion currents and toxicological screening work in intact embryos, as well as adult whole hearts, has demonstrated the utility of the zebrafish model to contribute to the development of therapeutics that lack hERG-blocking off-target effects. Moreover, forward and reverse genetic approaches show zebrafish as a tractable model in which LQTS2 can be studied. With the development of new tools and technologies, zebrafish lines carrying precise channel variants associated with LQTS2 have recently begun to be generated and explored. In this review, we discuss the present knowledge and questions raised related to the use of zebrafish as models of acquired and inherited LQTS2. We focus discussion, in particular, on developments in precise gene-editing approaches in zebrafish to create whole heart inherited LQTS2 models and evidence that zebrafish hearts can be used to study arrhythmogenicity and to identify potential anti-arrhythmic compounds.

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KHz-rate volumetric voltage imaging of the whole zebrafish heart

Cited by 4 publications

References 32 publications

Mapping volumes to planes: Camera-based strategies for snapshot volumetric microscopy

Mapping volumes to planes: Camera-based strategies for snapshot volumetric microscopy

Novel Optics-Based Approaches for Cardiac Electrophysiology: A Review

Utility of Zebrafish Models of Acquired and Inherited Long QT Syndrome

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