Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2020 Oct 30;10(11):886.
doi: 10.3390/diagnostics10110886.

Blood Vessel Imaging at Pre-Larval Stages of Zebrafish Embryonic Development

Alexander S Machikhin  1 , Mikhail V Volkov  2 , Alexander B Burlakov  3 , Demid D Khokhlov  1 , Andrey V Potemkin  2
Affiliations

Blood Vessel Imaging at Pre-Larval Stages of Zebrafish Embryonic Development

Alexander S Machikhin et al. Diagnostics (Basel). .

Abstract

The zebrafish (Danio rerio) is an increasingly popular animal model biological system. In cardiovascular research, it has been used to model specific cardiac phenomena as well as to identify novel therapies for human cardiovascular disease. While the zebrafish cardiovascular system functioning is well examined at larval stages, the mechanisms by which vessel activity is initiated remain a subject of intense investigation. In this research, we report on an in vivo stain-free blood vessel imaging technique at pre-larval stages of zebrafish embryonic development. We have developed the algorithm for the enhancement, alignment and spatiotemporal analysis of bright-field microscopy images of zebrafish embryos. It enables the detection, mapping and quantitative characterization of cardiac activity across the whole specimen. To validate the proposed approach, we have analyzed multiple data cubes, calculated vessel images and evaluated blood flow velocity and heart rate dynamics in the absence of any anesthesia. This non-invasive technique may shed light on the mechanism of vessel activity initiation and stabilization as well as the cardiovascular system's susceptibility to environmental stressors at early developmental stages.

Keywords: cardiovascular system; embryonic development; in vivo imaging; non-invasive measurements; optical mapping; zebrafish.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interests.

Figures

Figure 1
Figure 1
Experimental protocol.
Figure 2
Figure 2
Image processing pipeline.
Figure 3
Figure 3
Stages of image pre-processing: (a) cropped image, (b) aligned image, (c) locally matched image and (d) blood flow image.
Figure 4
Figure 4
Example of cardiovascular data extraction using the proposed algorithm: (a) blood flow images, (b) vessel image, (c) the selected heart area, (d) the selected vessel area, (e) offset vector field examples, (f) the detected vessel central line, (g) cardiac signal and blood flow velocity examples, (h) cardiac signal spectrum, (i) blood flow velocity range and average value.
Figure 5
Figure 5
Calculated vessel images (green rectangles indicate the detected heart area).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Kawasaki T., Maeno A., Shiroishi T., Sakai N. Development and growth of organs in living whole embryo and larval grafts in zebrafish. Sci. Rep. UK. 2017;7:16508. doi: 10.1038/s41598-017-16642-5. - DOI - PMC - PubMed
    1. Yalcin H.C., Amindari A., Butcher J.T., Althan A., Yacoub M. Heart function and hemodynamic analysis for zebrafish embryos. Dev. Dynam. 2017;246:868–880. doi: 10.1002/dvdy.24497. - DOI - PubMed
    1. Lawson N.D., Weinstein B.M. Arteries and veins: Making a difference with zebrafish. Nat. Rev. Genet. 2002;3:674–682. doi: 10.1038/nrg888. - DOI - PubMed
    1. Konno M., Asai A., Kitagawa T., Yabumoto M., Ofusa K., Arai T., Hirotsu T., Doki Y., Eguchi H., Ishii H. State-of-the-Art Technology of Model Organisms for Current Human Medicine. Diagnostics. 2020;10:392. doi: 10.3390/diagnostics10060392. - DOI - PMC - PubMed
    1. Soroldoni D., Hogan B.M., Oates A.C. Simple and efficient transgenesis with meganuclease constructs in zebrafish. Methods Mol. Biol. 2009;546:117–130. doi: 10.1007/978-1-60327-977-2_8. - DOI - PubMed

LinkOut - more resources