Four‐dimensional live imaging of hemodynamics in mammalian embryonic heart with Doppler optical coherence tomography

Hemodynamic analysis of the mouse embryonic heart is essential for understanding the functional aspects of early cardiogenesis and advancing the research in congenital heart defects. However, high-resolution imaging of cardiac hemodynamics in mammalian models remains challenging, primarily due to the dynamic nature and deep location of the embryonic heart. Here we report four-dimensional micro-scale imaging of blood flow in the early mouse embryonic heart, enabling time-resolved measurement and analysis of flow velocity throughout the heart tube. Our method uses Doppler optical coherence tomography in live mouse embryo culture, and employs a post-processing synchronization approach to reconstruct three-dimensional data over time at a 100 Hz volume rate. Experiments were performed on live mouse embryos at embryonic day 9.0. Our results show blood flow dynamics inside the beating heart, with the capability for quantitative flow velocity assessment in the primitive atrium, atrioventricular and bulboventricular regions, and bulbus cordis. Combined cardiodynamic and hemodynamic analysis indicates this functional imaging method can be utilized to further investigate the mechanical relationship between blood flow dynamics and cardiac wall movement, bringing new possibilities to study biomechanics in early mammalian cardiogenesis. Four-dimensional live hemodynamic imaging of the mouse embryonic heart at embryonic day 9.0 using Doppler optical coherence tomography, showing directional blood flows in the sinus venosus, primitive atrium, atrioventricular region and vitelline vein.

[1]  Michael W. Jenkins,et al.  Ultrahigh-speed optical coherence tomography imaging and visualization of the embryonic avian heart using a buffered Fourier Domain Mode Locked laser. , 2007, Optics express.

[2]  Michael W. Jenkins,et al.  4D embryonic cardiography using gated optical coherence tomography. , 2006, Optics express.

[3]  Kirill V Larin,et al.  Imaging mouse embryonic cardiovascular development. , 2012, Cold Spring Harbor protocols.

[4]  J. Nelson,et al.  Characterization of fluid flow velocity by optical Doppler tomography. , 1995, Optics letters.

[5]  J. Izatt,et al.  In vivo spectral domain optical coherence tomography volumetric imaging and spectral Doppler velocimetry of early stage embryonic chicken heart development. , 2008, Journal of the Optical Society of America. A, Optics, image science, and vision.

[6]  Alex Cable,et al.  Live imaging of blood flow in mammalian embryos using Doppler swept-source optical coherence tomography. , 2008, Journal of biomedical optics.

[7]  L A Taber,et al.  Mechanical aspects of cardiac development. , 1998, Progress in biophysics and molecular biology.

[8]  Julius Pekar,et al.  High speed, wide velocity dynamic range Doppler optical coherence tomography (Part I): System design, signal processing, and performance. , 2003, Optics express.

[9]  D. A. Christopher,et al.  40-MHZ echocardiography scanner for cardiovascular assessment of mouse embryos. , 1998, Ultrasound in medicine & biology.

[10]  Gabriel Acevedo-Bolton,et al.  Intracardiac fluid forces are an essential epigenetic factor for embryonic cardiogenesis , 2003, Nature.

[11]  Willy Supatto,et al.  Advances in multiphoton microscopy for imaging embryos. , 2011, Current opinion in genetics & development.

[12]  Robert G. Gourdie,et al.  Hemodynamics Is a Key Epigenetic Factor in Development of the Cardiac Conduction System , 2003, Circulation research.

[13]  Jörg Stypmann,et al.  Doppler Ultrasound in Mice , 2007, Echocardiography.

[14]  Daniel H Turnbull,et al.  Ultrasound biomicroscopy-Doppler in mouse cardiovascular development. , 2003, Physiological genomics.

[15]  Taner Akkin,et al.  Spectral-domain optical coherence phase microscopy for quantitative phase-contrast imaging. , 2005, Optics letters.

[16]  Kirill V. Larin,et al.  Direct four-dimensional structural and functional imaging of cardiovascular dynamics in mouse embryos with 1.5 MHz optical coherence tomography. , 2015, Optics letters.

[17]  Leopold Schmetterer,et al.  Doppler Optical Coherence Tomography , 2014, Progress in Retinal and Eye Research.

[18]  F. Chang,et al.  Systemic assessment of fetal hemodynamics by Doppler ultrasound. , 2000, Ultrasound in medicine & biology.

[19]  Ruikang K. Wang,et al.  Efficient postacquisition synchronization of 4-D nongated cardiac images obtained from optical coherence tomography: application to 4-D reconstruction of the chick embryonic heart. , 2009, Journal of biomedical optics.

[20]  Yixian Zheng,et al.  Lineage mapping the pre-implantation mouse embryo by two-photon microscopy, new insights into the segregation of cell fates. , 2011, Developmental biology.

[21]  David L Wilson,et al.  High temporal resolution OCT using image-based retrospective gating. , 2009, Optics express.

[22]  Lixin Chin,et al.  Three-dimensional optical coherence elastography by phase-sensitive comparison of C-scans , 2014, Journal of biomedical optics.

[23]  T. Mikawa,et al.  Hemodynamic-dependent patterning of endothelin converting enzyme 1 expression and differentiation of impulse-conducting Purkinje fibers in the embryonic heart , 2004, Development.

[24]  C S Peskin,et al.  Hemodynamics in congenital heart disease. , 1986, Computers in biology and medicine.

[25]  Kirill V Larin,et al.  Live four-dimensional optical coherence tomography reveals embryonic cardiac phenotype in mouse mutant. , 2015, Journal of biomedical optics.

[26]  John White,et al.  Long-term two-photon fluorescence imaging of mammalian embryos without compromising viability , 1999, Nature Biotechnology.

[27]  J. Freund,et al.  Blood flow mechanics in cardiovascular development , 2015, Cellular and Molecular Life Sciences.

[28]  D. Turnbull,et al.  Onset of Cardiac Function During Early Mouse Embryogenesis Coincides With Entry of Primitive Erythroblasts Into the Embryo Proper , 2003, Circulation research.

[29]  Michael Liebling,et al.  Rapid three‐dimensional imaging and analysis of the beating embryonic heart reveals functional changes during development , 2006, Developmental dynamics : an official publication of the American Association of Anatomists.

[30]  Joseph Izatt,et al.  Quantitative Measurement of Blood Flow Dynamics in Embryonic Vasculature Using Spectral Doppler Velocimetry , 2009, Anatomical record.

[31]  M. Dickinson,et al.  Hemodynamic measurements from individual blood cells in early mammalian embryos with Doppler swept source OCT. , 2009, Optics letters.

[32]  Michael Liebling,et al.  Reversing Blood Flows Act through klf2a to Ensure Normal Valvulogenesis in the Developing Heart , 2009, PLoS biology.

[33]  M E Dickinson,et al.  Dynamic in vivo imaging of postimplantation mammalian embryos using whole embryo culture , 2002, Genesis.

[34]  Jeffrey Robbins,et al.  Principles of genetic murine models for cardiac disease. , 2007, Circulation.

[35]  J. Izatt,et al.  High resolution imaging of in vivo cardiac dynamics using color Doppler optical coherence tomography. , 1997, Optics express.

[36]  K. Nicolaides,et al.  Assessment of fetal compromise by Doppler ultrasound investigation of the fetal circulation. Arterial, intracardiac, and venous blood flow velocity studies. , 1995, Circulation.

[37]  Andrew L. Lopez,et al.  Noncontact quantitative biomechanical characterization of cardiac muscle using shear wave imaging optical coherence tomography. , 2014, Biomedical optics express.

[38]  Norihiko Nishizawa,et al.  Quantitative comparison of contrast and imaging depth of ultrahigh-resolution optical coherence tomography images in 800–1700 nm wavelength region , 2012, Biomedical optics express.

[39]  Benjamin J Vakoc,et al.  Multimodality optical imaging of embryonic heart microstructure. , 2007, Journal of biomedical optics.

[40]  Brian C. Wilson,et al.  Improved phase-resolved optical Doppler tomography using the Kasai velocity estimator and histogram segmentation , 2002 .

[41]  Jay R Hove,et al.  Quantifying Cardiovascular Flow Dynamics During Early Development , 2006, Pediatric Research.

[42]  Adrian Bradu,et al.  Closed loop tracked Doppler optical coherence tomography based heart monitor for the Drosophila melanogaster larvae , 2016, Journal of biophotonics.

[43]  Daniel L Marks,et al.  Three-dimensional optical coherence tomography of the embryonic murine cardiovascular system. , 2006, Journal of biomedical optics.

[44]  Anne Moon,et al.  Mouse models of congenital cardiovascular disease. , 2008, Current topics in developmental biology.

[45]  S. Colan,et al.  Bulboventricular foramen size in infants with double-inlet left ventricle or tricuspid atresia with transposed great arteries: influence on initial palliative operation and rate of growth. , 1992, Journal of the American College of Cardiology.

[46]  Sonja Nowotschin,et al.  Imaging mouse development with confocal time-lapse microscopy. , 2010, Methods in enzymology.

[47]  Michael Liebling,et al.  Live Imaging of Early Developmental Processes in Mammalian Embryos with Optical Coherence Tomography. , 2009, Journal of innovative optical health sciences.

[48]  Irina V. Larina,et al.  Vascular development and hemodynamic force in the mouse yolk sac , 2014, Front. Physiol..

[49]  Joseph A. Izatt,et al.  Optical Coherence Tomography: A New High-Resolution Imaging Technology to Study Cardiac Development in Chick Embryos , 2002, Circulation.

[50]  Andrew M. Rollins,et al.  Capturing structure and function in an embryonic heart with biophotonic tools , 2014, Front. Physiol..

[51]  Venkat Keshav Chivukula,et al.  Blood flow through the embryonic heart outflow tract during cardiac looping in HH13–HH18 chicken embryos , 2015, Journal of The Royal Society Interface.

[52]  J. Freund,et al.  Fluid flows and forces in development: functions, features and biophysical principles , 2012, Development.

[53]  Scott E Fraser,et al.  Vascular remodeling of the mouse yolk sac requires hemodynamic force , 2007, Development.

[54]  Ruikang K. Wang,et al.  Biomechanics of the Chick Embryonic Heart Outflow Tract at HH18 Using 4D Optical Coherence Tomography Imaging and Computational Modeling , 2012, PloS one.

[55]  J. Hoffman,et al.  The incidence of congenital heart disease. , 2002, Journal of the American College of Cardiology.

[56]  A. Becker,et al.  Morphogenesis of univentricular hearts. , 1976, British heart journal.

[57]  M E Dickinson,et al.  Measuring hemodynamic changes during mammalian development. , 2004, American journal of physiology. Heart and circulatory physiology.

[58]  Michael Liebling,et al.  Four-dimensional cardiac imaging in living embryos via postacquisition synchronization of nongated slice sequences. , 2005, Journal of biomedical optics.

[59]  B. Furst Hemodynamics of the Early Embryo Circulation , 2019, The Heart and Circulation.

[60]  David S. Koos,et al.  Deep and fast live imaging with two-photon scanned light-sheet microscopy , 2011, Nature Methods.

[61]  Adrian Mariampillai,et al.  Doppler optical cardiogram gated 2D color flow imaging at 1000 fps and 4D in vivo visualization of embryonic heart at 45 fps on a swept source OCT system. , 2007, Optics express.

[62]  Aneeka M Hancock,et al.  Fully distributed absolute blood flow velocity measurement for middle cerebral arteries using Doppler optical coherence tomography. , 2016, Biomedical optics express.

[63]  Shang Wang,et al.  Optical coherence tomography guided microinjections in live mouse embryos: high-resolution targeted manipulation for mouse embryonic research , 2015, Journal of biomedical optics.

[64]  Irina V Larina,et al.  A Membrane Associated mCherry Fluorescent Reporter Line for Studying Vascular Remodeling and Cardiac Function During Murine Embryonic Development , 2009, Anatomical record.

[65]  David L Wilson,et al.  Measuring hemodynamics in the developing heart tube with four-dimensional gated Doppler optical coherence tomography. , 2010, Journal of biomedical optics.