An Optimized Method for Delivering Flow Tracer Particles to Intravital Fluid Environments in the Developing Zebrafish

Craig, Michael P.; Gilday, Steven D.; Dabiri, Dana; Hove, Jay R.

doi:10.1089/zeb.2012.0740

Cited by 21 publications

(14 citation statements)

References 45 publications

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“…The total blood volume of zebrafish embryos at 2 dpf has been estimated to be 60 nL (Craig et al, 2012), so the blood volume is not significantly altered upon injection. Immediately after the injections, larvae were embedded in 1.2% low-melting-point agarose (Invitrogen) made in egg water.…”

Section: Methodsmentioning

confidence: 99%

Functional and genetic analysis of choroid plexus development in zebrafish

Henson

Parupalli

et al. 2014

Front. Neurosci.

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The choroid plexus, an epithelial-based structure localized in the brain ventricle, is the major component of the blood-cerebrospinal fluid barrier. The choroid plexus produces the cerebrospinal fluid and regulates the components of the cerebrospinal fluid. Abnormal choroid plexus function is associated with neurodegenerative diseases, tumor formation in the choroid plexus epithelium, and hydrocephaly. In this study, we used zebrafish (Danio rerio) as a model system to understand the genetic components of choroid plexus development. We generated an enhancer trap line, Et(cp:EGFP)sj2, that expresses enhanced green fluorescent protein (EGFP) in the choroid plexus epithelium. Using immunohistochemistry and fluorescent tracers, we demonstrated that the zebrafish choroid plexus possesses brain barrier properties such as tight junctions and transporter activity. Thus, we have established zebrafish as a functionally relevant model to study choroid plexus development. Using an unbiased approach, we performed a forward genetic dissection of the choroid plexus to identify genes essential for its formation and function. Using Et(cp:EGFP)sj2, we isolated 10 recessive mutant lines with choroid plexus abnormalities, which were grouped into five classes based on GFP intensity, epithelial localization, and overall choroid plexus morphology. We also mapped the mutation for two mutant lines to chromosomes 4 and 21, respectively. The mutants generated in this study can be used to elucidate specific genes and signaling pathways essential for choroid plexus development, function, and/or maintenance and will provide important insights into how these genetic mutations contribute to disease.

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Section: Methodsmentioning

confidence: 99%

Functional and genetic analysis of choroid plexus development in zebrafish

Henson

Parupalli

et al. 2014

Front. Neurosci.

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“…Despite the sub-optimal spatial resolution for the densely organized cardiomyocytes, LFM demonstrated the capability for tracking the sparsely distributed signals like blood cells (Truong et al, 2020;Wagner et al, 2019;Wang et al, 2020). This is further applicable to study the myocardial Calcium (Ca 2+ ) flux via the genetically encoded Ca 2+ indicators (GECIs) such as GCaMP for electromechanical coupling (Weber et al, 2017) and particle tracers such as nanoparticles (Craig et al, 2012) to extend the application of our method.…”

Section: Integration Of Light-field and Light-sheet To Capture Myocarmentioning

confidence: 94%

A Hybrid of Light-Field and Light-Sheet Imaging to Decouple Myocardial Biomechanics from Intracardiac Flow Dynamics

Wang¹,

Ding²,

Satta³

et al. 2020

Preprint

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Biomechanical forces intimately contribute to cardiac morphogenesis. However, 4-D (3-D space + time) imaging is needed to investigate the developmental cardiac mechanics with high temporal and spatial resolution. We hereby integrated light-sheet fluorescence microscopy (LSFM) with light-field microscopy (LFM), to simultaneously visualize myocardial contractility and intracardiac blood flow in three dimensions at 200 volumes per second (vps). LSFM allows for reconstruction of the myocardial contraction in zebrafish embryo; and LFM enables simultaneous tracking of the blood cells entering and leaving the contracting heart. We herein established particle tracking velocimetry to interrogate the trajectories of intracardiac blood cells, and we demonstrated deformable image registration to reveal a decrease in the myocardial contractility from atrioventricular (AV) canal to the outflow tract (OFT). We imaged myocardium undergoing torsional contraction and blood flow undergoing regurgitation. Taken together, the integration of light-field and light-sheet microscopy, followed by an image-based analysis pipeline, provides the biomechanical insights into coupling myocardial kinetics with rotational contraction along with intracardiac flow dynamics during development.

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“…Tracers providing high contrast are crucial to yield precise measurement of the blood flow [41]. We employ fluorescent beads as tracers since they are bright and uniform in size and shape, making the particle tracking more robust than the use of endogenous tracers such as the blood cells.…”

Section: Methodsmentioning

confidence: 99%

High-speed extended-volume blood flow measurement using engineered point-spread function

Zhou¹,

Žičkus²,

Zammit³

et al. 2018

Biomed. Opt. Express

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Experimental characterization of blood flow in living organisms is crucial for understanding the development and function of cardiovascular systems, but there has been no technique reported for snapshot imaging of thick samples in large volumes with high precision. We have combined computational microscopy and the diffraction-free, self-bending property of Airy-beams to track fluorescent beads with sub-micron precision through an extended axial range (up to 600 μm) within the flowing blood of 3 days post-fertilization (dpf) zebrafish embryos. The spatial trajectories of the tracer beads within flowing blood were recorded during transit through both cardinal and intersegmental vessels, and the trajectories were found to be consistent with the segmentation of the vasculature recorded using selective-plane illumination microscopy (SPIM). This method provides sufficiently precise spatial and temporal measurement of 3D blood flow that has the potential for directly probing key biomechanical quantities such as wall shear stress, as well as exploring the fluidic repercussions of cardiovascular diseases. Although we demonstrate the technique for blood flow, the ten-fold better enhancement in the depth range offers improvements in a wide range of applications of high-speed precision measurement of fluid flow, from microfluidics through measurement of cell dynamics to macroscopic aerosol characterizations.

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An Optimized Method for Delivering Flow Tracer Particles to Intravital Fluid Environments in the Developing Zebrafish

Cited by 21 publications

References 45 publications

Functional and genetic analysis of choroid plexus development in zebrafish

Functional and genetic analysis of choroid plexus development in zebrafish

A Hybrid of Light-Field and Light-Sheet Imaging to Decouple Myocardial Biomechanics from Intracardiac Flow Dynamics

High-speed extended-volume blood flow measurement using engineered point-spread function

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