A Stretching Device for High-Resolution Live-Cell Imaging

Huang, Lawrence; Mathieu, Pattie S.; Helmke, Brian P.

doi:10.1007/s10439-010-9968-7

Cited by 69 publications

(69 citation statements)

References 27 publications

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“…This is the first example of a strain device that can be used to assess biological function at the tissue level. All previous devices that have been designed to view high-resolution images of mechanically strained samples have been done at the cell level (Huang et al 2010;Gerstmair et al 2009;Wang et al 2010). In essence, these systems require isolated cells to be attached to an elastic membrane before straining.…”

Section: Discussionmentioning

confidence: 99%

Live en face imaging of aortic valve leaflets under mechanical stress

Metzler

Digesu

Howard

et al. 2011

Biomech Model Mechanobiol

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Soft tissues, such as tendons, skin, arteries, or lung, are constantly subject to mechanical stresses in vivo. None more so than the aortic heart valve that experiences an array of forces including shear stress, cyclic pressure, strain, and flexion. Anisotropic biaxial cyclic stretch maintains valve homeostasis; however, abnormal forces are implicated in disease progression. The response of the valve endothelium to deviations from physiological levels has not been fully characterized. Here, we show the design and validation of a novel stretch apparatus capable of applying biaxial stretch to viable heart valve tissue, while simultaneously allowing for live en face endothelial cell imaging via confocal laser scanning microscopy (CLSM). Real-time imaging of tissue is possible while undergoing highly characterized mechanical conditions and maintaining the native extracellular matrix. Thus, it provides significant advantages over traditional cell culture or in vivo animal models. Planar biaxial tissue stretching with simultaneous live cell imaging could prove useful in studying the mechanobiology of any soft tissue.

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Section: Discussionmentioning

confidence: 99%

Live en face imaging of aortic valve leaflets under mechanical stress

Metzler

Digesu

Howard

et al. 2011

Biomech Model Mechanobiol

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“…Another important modification is live-cell visualization, which can open up new possibilities for relevant data collection during mechanostimulation studies [107,108]. Several systems have been developed to facilitate live-cell imaging in uniaxial strain systems [95,106,108]. These may include a compensation motor to keep the same cell region within the field of view [108].…”

Section: Strain Methodsmentioning

confidence: 99%

“…With any stretch modality (uniaxial, equiaxial, or biaxial), validation of strain is important to determine whether cells in different regions are receiving a consistent stimulus. This may be accomplished by video tracking of visible markers or fluorescent dots on the substrate during stretch, sometimes in conjunction with a displacement transducer [91][92][93][94][95][96][97][98][99]. Tension on the membrane can be measured using a load cell [96].…”

Section: Strain Methodsmentioning

confidence: 99%

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Device-Based In Vitro Techniques for Mechanical Stimulation of Vascular Cells: A Review

Davis

Zambrano

Anumolu

et al. 2015

Journal of Biomechanical Engineering

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The most common cause of death in the developed world is cardiovascular disease. For decades, this has provided a powerful motivation to study the effects of mechanical forces on vascular cells in a controlled setting, since these cells have been implicated in the development of disease. Early efforts in the 1970 s included the first use of a parallel-plate flow system to apply shear stress to endothelial cells (ECs) and the development of uniaxial substrate stretching techniques (Krueger et al., 1971, "An in Vitro Study of Flow Response by Cells," J. Biomech., 4(1), pp. 31-36 and Meikle et al., 1979, "Rabbit Cranial Sutures in Vitro: A New Experimental Model for Studying the Response of Fibrous Joints to Mechanical Stress," Calcif. Tissue Int., 28(2), pp. 13-144). Since then, a multitude of in vitro devices have been designed and developed for mechanical stimulation of vascular cells and tissues in an effort to better understand their response to in vivo physiologic mechanical conditions. This article reviews the functional attributes of mechanical bioreactors developed in the 21st century, including their major advantages and disadvantages. Each of these systems has been categorized in terms of their primary loading modality: fluid shear stress (FSS), substrate distention, combined distention and fluid shear, or other applied forces. The goal of this article is to provide researchers with a survey of useful methodologies that can be adapted to studies in this area, and to clarify future possibilities for improved research methods.

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“…et Hornberger et al, 2005), or characterization of cellular (Gavara et al, 2008;Huang et al, 2010) and nuclear (Lammerding et al, 2007) deformations. However, these methods are restricted to applications in 2D cell culture systems.…”

Section: Introductionmentioning

confidence: 99%

Quantification of substrate and cellular strains in stretchable 3D cell cultures: an experimental and computational framework

Gonzalez-Avalos

Mürnseer

Deeg

et al. 2017

Journal of Microscopy

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SummaryThe mechanical cell environment is a key regulator of biological processes . In living tissues, cells are embedded into the 3D extracellular matrix and permanently exposed to mechanical forces. Quantification of the cellular strain state in a 3D matrix is therefore the first step towards understanding how physical cues determine single cell and multicellular behaviour. The majority of cell assays are, however, based on 2D cell cultures that lack many essential features of the in vivo cellular environment. Furthermore, nondestructive measurement of substrate and cellular mechanics requires appropriate computational tools for microscopic image analysis and interpretation. Here, we present an experimental and computational framework for generation and quantification of the cellular strain state in 3D cell cultures using a combination of 3D substrate stretcher, multichannel microscopic imaging and computational image analysis. The 3D substrate stretcher enables deformation of living cells embedded in bead-labelled 3D collagen hydrogels. Local substrate and cell deformations are determined by tracking displacement of fluorescent beads with subsequent finite element interpolation of cell strains over a tetrahedral tessellation. In this feasibility study, we debate diverse aspects of deformable 3D culture construction, ¶ These two authors contribute equally.

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A Stretching Device for High-Resolution Live-Cell Imaging

Cited by 69 publications

References 27 publications

Live en face imaging of aortic valve leaflets under mechanical stress

Live en face imaging of aortic valve leaflets under mechanical stress

Device-Based In Vitro Techniques for Mechanical Stimulation of Vascular Cells: A Review

Quantification of substrate and cellular strains in stretchable 3D cell cultures: an experimental and computational framework

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