Live imaging of blood flow in mammalian embryos using Doppler swept-source optical coherence tomography

Larina, Irina V.; Sudheendran, Narendran; Ghosn, Mohamad G.; Jiang, James; Cable, Alex; Larin, Kirill V.; Dickinson, Mary E.

doi:10.1117/1.3046716

Cited by 97 publications

(91 citation statements)

References 16 publications

(13 reference statements)

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“…Some studies have successfully performed live imaging of blood flow in ex vivo murine embryos using Doppler swept-source optical coherence tomography 39,40 , while magnetic resonance imaging, vascular corrosion casts, microcomputed tomography and optical projection tomography have been implemented postmortem 16 . This is unfortunate, since this period of embryonic growth can reveal crucial information regarding early vascular development and function.…”

Section: Discussionmentioning

confidence: 99%

Contrast Imaging in Mouse Embryos Using High-frequency Ultrasound

Denbeigh

Nixon

Puri

et al. 2015

JoVE

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Ultrasound contrast-enhanced imaging can convey essential quantitative information regarding tissue vascularity and perfusion and, in targeted applications, facilitate the detection and measure of vascular biomarkers at the molecular level. Within the mouse embryo, this noninvasive technique may be used to uncover basic mechanisms underlying vascular development in the early mouse circulatory system and in genetic models of cardiovascular disease. The mouse embryo also presents as an excellent model for studying the adhesion of microbubbles to angiogenic targets (including vascular endothelial growth factor receptor 2 (VEGFR2) or α v β 3 ) and for assessing the quantitative nature of molecular ultrasound. We therefore developed a method to introduce ultrasound contrast agents into the vasculature of living, isolated embryos. This allows freedom in terms of injection control and positioning, reproducibility of the imaging plane without obstruction and motion, and simplified image analysis and quantification. Late gestational stage (embryonic day (E)16.6 and E17.5) murine embryos were isolated from the uterus, gently exteriorized from the yolk sac and microbubble contrast agents were injected into veins accessible on the chorionic surface of the placental disc. Nonlinear contrast ultrasound imaging was then employed to collect a number of basic perfusion parameters (peak enhancement, wash-in rate and time to peak) and quantify targeted microbubble binding in an endoglin mouse model. We show the successful circulation of microbubbles within living embryos and the utility of this approach in characterizing embryonic vasculature and microbubble behavior. Video LinkThe video component of this article can be found at

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Section: Discussionmentioning

confidence: 99%

Contrast Imaging in Mouse Embryos Using High-frequency Ultrasound

Denbeigh

Nixon

Puri

et al. 2015

JoVE

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“…The embryos were given at least 30 mins to recover from the extraction procedure in a 37°C incubator with 5% CO 2 [37]. After the recovery period, the embryos were imaged, also while immersed in the DMEM culture solution.…”

Section: Embryo Preparation and Imagingmentioning

confidence: 99%

“…The sample arm of the OCT system was placed in the incubator to ensure viability of the embryos during live imaging. The images were corrected to physical dimensions assuming that the refractive index of the media and embryos was 1.4 [37]. …”

Section: Optical Coherence Tomography (Oct) Systemmentioning

confidence: 99%

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Applicability, usability, and limitations of murine embryonic imaging with optical coherence tomography and optical projection tomography

Singh¹,

Raghunathan²,

Piazza³

et al. 2016

Biomed. Opt. Express

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Abstract:We present an analysis of imaging murine embryos at various embryonic developmental stages (embryonic day 9.5, 11.5, and 13.5) by optical coherence tomography (OCT) and optical projection tomography (OPT). We demonstrate that while OCT was capable of rapid highresolution live 3D imaging, its limited penetration depth prevented visualization of deeper structures, particularly in later stage embryos. In contrast, OPT was able to image the whole embryos, but could not be used in vivo because the embryos must be fixed and cleared. Moreover, the fixation process significantly altered the embryo morphology, which was quantified by the volume of the eye-globes before and after fixation. All of these factors should be weighed when determining which imaging modality one should use to achieve particular goals of a study. References and links1. J. Beckers, W. Wurst, and M. H. de Angelis, "Towards better mouse models: enhanced genotypes, systemic phenotyping and envirotype modelling," Nat. Rev. Genet. 10(6), 371-380 (2009 440-443 (2000). 4. K. Paigen and J. T. Eppig, "A mouse phenome project," Mamm. Genome 11(9), 715-717 (2000). Proc. Natl. Acad. Sci. U.S.A. 91(9), 3530-3533 (1994). 9. M. Dhenain, S. W. Ruffins, and R. E. Jacobs, "Three-dimensional digital mouse atlas using high-resolution MRI," Dev. Biol. 232(2), 458-470 (2001 95-105 (1999). 33. U. Morgner, W. Drexler, F. X. Kärtner, X. D. Li, C. Pitris, E. P. Ippen, and J. G. Fujimoto, "Spectroscopic optical coherence tomography," Opt. Lett. 25(2), 111-113 (2000). 34. S. Rugonyi, C. Shaut, A. Liu, K. Thornburg, and R. K. Wang, "Changes in wall motion and blood flow in the outflow tract of chick embryonic hearts observed with optical coherence tomography after outflow tract banding and vitelline-vein ligation," Phys.

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“…Progress in OCT technology, for instance functional imaging [248,249], high-speed data acquisition [244] or advanced signal processing [250], has swiftly been adopted in developmental biology. These enabled further studies in early heart development and blood flow in various models establishing OCT as an important imaging tool for basic research.…”

Section: Developmental Biologymentioning

confidence: 99%

Optical coherence tomography—current technology and applications in clinical and biomedical research

et al. 2011

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Optical coherence tomography (OCT) is a non-invasive imaging technique providing real-time twoand three-dimensional images of scattering samples with micrometer resolution. Mapping the local reflectivity, OCT visualizes the morphology of the sample. In addition, functional properties such as birefringence, motion or the distribution of certain substances can be detected with high spatial resolution. The main field of application is bio-medical imaging and diagnostics. In ophthalmology, OCT is accepted as a clinical standard for diagnosing and monitoring treatment of a number of retinal diseases, and OCT is becoming an important instrument for clinical cardiology. New applications are emerging in various medical fields, e. g. early-stage cancer detection, surgical guidance, and early diagnosis of musculoskeletal diseases. OCT has also proven its value as a tool for developmental biology. The number of companies involved in manufacturing OCT systems has increased substantially during the last years-especially due to its success in opthalmology-and this technology can be expected to continue to spread into various fields of application.

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Live imaging of blood flow in mammalian embryos using Doppler swept-source optical coherence tomography

Cited by 97 publications

References 16 publications

Contrast Imaging in Mouse Embryos Using High-frequency Ultrasound

Contrast Imaging in Mouse Embryos Using High-frequency Ultrasound

Applicability, usability, and limitations of murine embryonic imaging with optical coherence tomography and optical projection tomography

Optical coherence tomography—current technology and applications in clinical and biomedical research

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