Solid Phase Synthesis of a Functionalized Bis-Peptide Using …
Published 5/15/2012
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In JoVE (1)
Other Publications (5)
- Anatomical Record (Hoboken, N.J. : 2007)
- Journal of Biomechanical Engineering
- Journal of Biomechanical Engineering
- Conference Proceedings : ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Conference
- Annals of Biomedical Engineering
Articles by Benjamen A. Filas in JoVE
Tracking Morphogenetic Tissue Deformations in the Early Chick Embryo
1Department of Biomedical Engineering, Washington University, 2Institute for Information Transmission Problems, Russian Academy of Sciences, 3Department of Mechanical Engineering and Materials Science, Washington University
This article describes surface labeling and ex ovo tissue culture in the early chick embryo. Techniques amenable to time-lapse bright field, fluorescence, and optical coherence tomography imaging are presented. Tracking surface labels with high spatiotemporal resolution enables kinematic quantities such as morphogenetic strains (deformations) to be calculated in both two and three dimensions.
Other articles by Benjamen A. Filas on PubMed
Optical Coherence Tomography As a Tool for Measuring Morphogenetic Deformation of the Looping Heart
Optical coherence tomography (OCT) was used to investigate morphogenesis of the embryonic chick heart during the first phase of looping (c-looping), as the heart bends and twists into a c-shaped tube. The present study focuses on the morphomechanical effects of the splanchnopleure (SPL), a membrane that has been shown to play a major role in cardiac torsion by pressing against the ventral surface of the heart. Without the SPL, rightward torsion (rotation) is delayed. The images show that compressive forces exerted by the SPL alter the shapes of the heart tube and primitive atria, as well as their spatial relationships. The SPL normally holds the heart in the plane of the embryo and forces cardiac jelly (CJ) out of adjacent regions in the atria. When the SPL is removed, cross-sections become more circular, CJ is more uniformly distributed, and the heart displaces ventrally. In addition, OCT-based morphogenetic strain maps were measured during looping by tracking the three-dimensional motions of microspheres placed on the myocardium. The spatial-temporal patterns of the strains correlated well with the observed behavior of the heart, including delayed torsion that occurs in SPL-lacking embryos. These results illustrate the potential of OCT as a tool in studies of morphogenesis, as well as provide a better understanding of the mechanical forces that drive cardiac looping.
A New Method for Measuring Deformation of Folding Surfaces During Morphogenesis
During morphogenesis, epithelia (cell sheets) undergo complex deformations as they stretch, bend, and twist to form the embryo. Often these changes in shape create multivalued surfaces that can be problematic for strain measurements. This paper presents a method for quantifying deformation of such surfaces. The method requires four-dimensional spatiotemporal coordinates of a finite number of surface markers, acquired using standard imaging techniques. From the coordinates of the markers, various deformation measures are computed as functions of time and space using straightforward matrix algebra. This method accommodates sparse randomly scattered marker arrays, with reasonable errors in marker locations. The accuracy of the method is examined for some sample problems with exact solutions. Then, the utility of the method is illustrated by using it to measure surface stretch ratios and shear in the looping heart and developing brain of the early chick embryo. In these examples, microspheres are tracked using optical coherence tomography. This technique provides a new tool that can be used in studies of the mechanics of morphogenesis.
On Modeling Morphogenesis of the Looping Heart Following Mechanical Perturbations
Looping is a crucial early phase during heart development, as the initially straight heart tube (HT) deforms into a curved tube to lay out the basic plan of the mature heart. This paper focuses on the first phase of looping, called c-looping, when the HT bends ventrally and twists dextrally (rightward) to create a c-shaped tube. Previous research has shown that bending is an intrinsic process, while dextral torsion is likely caused by external forces acting on the heart. However, the specific mechanisms that drive and regulate looping are not yet completely understood. Here, we present new experimental data and finite element models to help define these mechanisms for the torsional component of c-looping. First, with regions of growth and contraction specified according to experiments on chick embryos, a three-dimensional model exhibits morphogenetic deformation consistent with observations for normal looping. Next, the model is tested further using experiments in which looping is perturbed by removing structures that exert forces on the heart--a membrane (splanchnopleure (SPL)) that presses against the ventral surface of the heart and the left and right primitive atria. In all cases, the model predicts the correct qualitative behavior. Finally, a two-dimensional model of the HT cross section is used to study a feedback mechanism for stress-based regulation of looping. The model is tested using experiments in which the SPL is removed before, during, and after c-looping. In each simulation, the model predicts the correct response. Hence, these models provide new insight into the mechanical mechanisms that drive and regulate cardiac looping.
Structured Light Imaging of Epicardial Mechanics
There is a need for accurate measurements of mechanical strain and motion of the heart both in vitro and in vivo. We have developed a new structured-light imaging system capable of epicardial shape measurement at 333 fps at a resolution of 768 × 768 pixels. Here we present proof-of-concept data from our system applied to a beating rabbit heart in vitro to measure epicardial mechanics. This method will allow high resolution mapping of epicardial strain and virtual immobilization of the heart for removal of motion artifacts from epicardial recordings with fluorescence dyes. This will allow mapping of transmembrane potential and calcium transients in a beating heart, including in vivo.
Mechanical Stress As a Regulator of Cytoskeletal Contractility and Nuclear Shape in Embryonic Epithelia
The mechano-sensitive responses of the heart and brain were examined in the chick embryo during Hamburger and Hamilton stages 10-12. During these early stages of development, cells in these structures are organized into epithelia. Isolated hearts and brains were compressed by controlled amounts of surface tension (ST) at the surface of the sample, and microindentation was used to measure tissue stiffness following several hours of culture. The response of both organs was qualitatively similar, as they stiffened under reduced loading. With increased loading, however, the brain softened while heart stiffness was similar to controls. In the brain, changes in nuclear shape and morphology correlated with these responses, as nuclei became more elliptical with decreased loading and rounder with increased loading. Exposure to the myosin inhibitor blebbistatin indicated that these changes in stiffness and nuclear shape are likely caused by altered cytoskeletal contraction. Computational modeling suggests that this behavior tends to return peak tissue stress back toward the levels it has in the intact heart and brain. These results suggest that developing cardiac and neural epithelia respond similarly to changes in applied loads by altering contractility in ways that tend to restore the original mechanical stress state. Hence, this study supports the view that stress-based mechanical feedback plays a role in regulating epithelial development.