Quantitative measurement of blood velocity in zebrafish with optical vector field tomography

Fieramonti, Luca; Foglia, Efrem; Malavasi, Stefano; D’Andrea, Cosimo; Valentini, Gianluca; Cotelli, Franco; Bassi, Andrea

doi:10.1002/jbio.201300162

Cited by 24 publications

(25 citation statements)

References 20 publications

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“…1a at 3.5 s, see also Supplementary Movie 3 ). Subsequently, the particles could be displaced and also moved against the direction of fast blood flow (200 μm s −1 in the vein and ∼700 μm s −1 in the artery 14 ) indicating strong trapping. In the example shown ( Fig.…”

Section: Resultsmentioning

confidence: 99%

Optical micromanipulation of nanoparticles and cells inside living zebrafish

et al. 2016

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Regulation of biological processes is often based on physical interactions between cells and their microenvironment. To unravel how and where interactions occur, micromanipulation methods can be used that offer high-precision control over the duration, position and magnitude of interactions. However, lacking an in vivo system, micromanipulation has generally been done with cells in vitro, which may not reflect the complex in vivo situation inside multicellular organisms. Here using optical tweezers we demonstrate micromanipulation throughout the transparent zebrafish embryo. We show that different cells, as well as injected nanoparticles and bacteria can be trapped and that adhesion properties and membrane deformation of endothelium and macrophages can be analysed. This non-invasive micromanipulation inside a whole-organism gives direct insights into cell interactions that are not accessible using existing approaches. Potential applications include screening of nanoparticle-cell interactions for cancer therapy or tissue invasion studies in cancer and infection biology.

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Section: Resultsmentioning

confidence: 99%

Optical micromanipulation of nanoparticles and cells inside living zebrafish

et al. 2016

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“…The increase of the efficiency of the nonlinear excitation obtained by the optical aberrations correction, the accuracy in the laser spot positioning, and the use of no movable parts make our setup particularly suited to characterize flows in complex vessel networks, 57,58 in confined environments, 59 or in vivo 8 by means of cross-correlation methods. For these applications, it is essential to be able to select the observation volumes on an image collected with the same setting of the optical setup, particularly for low brightness and signal/noise ratio samples.…”

Section: Aberrations Correction and Cross-correlation Spectroscopymentioning

confidence: 99%

Adaptive optics microspectrometer for cross-correlation measurement of microfluidic flows

et al. 2019

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Mapping flows in vivo is essential for the investigation of cardiovascular pathologies in animal models.The limitation of optical-based methods, such as space-time cross correlation, is the scattering of light by the connective and fat components and the direct wave front distortion by large inhomogeneities in the tissue. Nonlinear excitation of the sample fluorescence helps us by reducing light scattering in excitation. However, there is still a limitation on the signal-background due to the wave front distortion. We develop a diffractive optical microscope based on a single spatial light modulator (SLM) with no movable parts. We combine the correction of wave front distortions to the cross-correlation analysis of the flow dynamics. We use the SLM to shine arbitrary patterns of spots on the sample, to correct their optical aberrations, to shift the aberration corrected spot array on the sample for the collection of fluorescence images, and to measure flow velocities from the cross-correlation functions computed between couples of spots. The setup and the algorithms are tested on various microfluidic devices. By applying the adaptive optics correction algorithm, it is possible to increase up to 5 times the signalto-background ratio and to reduce approximately of the same ratio the uncertainty of the flow speed measurement. By working on grids of spots, we can correct different aberrations in different portions of the field of view, a feature that allows for anisoplanatic aberrations correction. Finally, being more efficient in the excitation, we increase the accuracy of the speed measurement by employing a larger number of spots in the grid despite the fact that the two-photon excitation efficiency scales as the fourth power of this number: we achieve a twofold decrease of the uncertainty and a threefold increase of the accuracy in the evaluation of the flow speed. © The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

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“…7 This has led to the discovery of vascular guidance and networking molecules as well as the identification of vessel maturation-regulating molecules, such as the angiopoietins and the platelet-derived growth factors. 8 In this field, medical research often exploits small animal models such as zebrafish embryos 9 and juvenile transparent fishes 10,11 for pilot in vivo studies. In addition, the advances of microfluidics are greatly influencing several fields of microbiology, providing new tools to investigate processes developing under flow, such as bacterial biofilm formation.…”

Section: Spatiotemporal Image Correlation Analysis Of Blood Flow In Bmentioning

confidence: 99%

Spatiotemporal image correlation analysis of blood flow in branched vessel networks of zebrafish embryos

et al. 2017

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Ramification of blood circulation is relevant in a number of physiological and pathological conditions. The oxygen exchange occurs largely in the capillary bed, and the cancer progression is closely linked to the angiogenesis around the tumor mass. Optical microscopy has made impressive improvements in in vivo imaging and dynamic studies based on correlation analysis of time stacks of images. Here, we develop and test advanced methods that allow mapping the flow fields in branched vessel networks at the resolution of 10 to 20 μm. The methods, based on the application of spatiotemporal image correlation spectroscopy and its extension to cross-correlation analysis, are applied here to the case of early stage embryos of zebrafish.

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Quantitative measurement of blood velocity in zebrafish with optical vector field tomography

Cited by 24 publications

References 20 publications

Optical micromanipulation of nanoparticles and cells inside living zebrafish

Optical micromanipulation of nanoparticles and cells inside living zebrafish

Adaptive optics microspectrometer for cross-correlation measurement of microfluidic flows

Spatiotemporal image correlation analysis of blood flow in branched vessel networks of zebrafish embryos

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