China
Africa
Explore chapters and articles related to this topic
Computational and Experimental Approaches to Cellular and Subcellular Tracking at the Nanoscale
Published in Sarhan M. Musa, ®, 2018
Zeinab Al-Rekabi, Dominique Tremblay, Kristina Haase, Richard L. Leask, Andrew E. Pelling
Cells are also sensitive to mechanical properties of the microenvironment in which they find themselves. Specifically it has been demonstrated that the stiffness, or Young’s Modulus, of the substrate on which cells are growing can dictate the stem cell fate (Engler et al. 2006), the formation of new muscle fibers (Engler et al. 2004; Discher et al. 2005; McDaniel et al. 2007; Zhang et al. 2009), the progression of apoptosis (Wang et al. 2000; Pelling et al. 2009) and cell adherence, migration, and proliferation (Dembo and Wang 1999; Lo et al. 2000; Bershadsky et al. 2003). Many of these cellular responses to the mechanical microenvironment are driven by the ability of cells to pull and push on their substrate. These cellular forces are known as cellular traction forces (CTFs) and are driven in large part by the underlying architecture of the cell. The feedback between the stiffness of the substrate and the magnitude of the forces the cell exert have profound impacts on cellular behavior (Lo et al. 2000; Balaban et al. 2001; Beningo et al. 2001; Munevar et al. 2001; Cai and Sheetz 2009). Therefore, measuring and quantifying CTF have important biomedical relevance. In a technique known as traction force microscopy (TFM) (Dembo and Wang 1999; Lo et al. 2000; Munevar et al. 2001; Beningo et al. 2006), cells are cultured on flexible polymer substrates in which nanoscale fluorescent beads have been embedded as fiduciary markers. As cells exert “pulling” and “pushing” forces on the flexible substrate, the marker beads move proportionately. Computational image routines that are capable of tracking fiduciary marker displacements allow one to determine the magnitude of the exerted forces (Dembo and Wang 1999).
Imaging Cell Adhesion and Migration
Published in Margarida M. Barroso, Xavier Intes, In Vivo, 2020
Chandrani Mondal, Julie Di Martino, Jose Javier Bravo-Cordero
Other imaging tools to study focal adhesion biology include traction force microscopy, atomic force microscopy (AFM), AFM-based force spectroscopy, fluorescence microscopy and fluid force microscopy-based single-cell force spectroscopy, and photonic crystal enhanced microscopy (PCEM). Traction force microscopy is a technique that is particularly useful for measuring cell traction forces in generating mechanical signals driving adhesion dynamics and allows for the analysis of both adhesion molecule dynamics and ECM deformation by the cell contact (Wang and Li, 2009). Atomic force microscopy (AFM) for example, was used in a study in which the adhesion forces of cervical carcinoma cells in tissue culture were measured by using the manipulation force microscope, a novel AFM (Sagvolden et al., 1999). AFM-based force spectroscopy has been used to quantify single-molecule adhesion forces in living amoeboid cells (Eibl and Benoit, 2004). Kashef and Franz (2015) used a combination of TIRF and AFM to study adhesion and to better understand the initial adhesion events when cells engage contact with the ECM. Sankaran et al. (2017) used fluorescence microscopy and fluid force microscopy-based single-cell force spectroscopy to show that actin filaments, focal adhesions, adhesion forces, and cell contractility are comparable between cells adhering to covalent and noncovalent surfaces. Photonic crystal enhanced microscopy (PCEM) is a novel biosensor-based microscopy technique that allows for the movement of cellular materials at the plasma membrane of individual living cells to be monitored dynamically and imaged quantitatively (Zhuo et al., 2016). This is a high sensitivity label-free live-cell imaging technique allowing for the profiling of dynamic adhesions of single cells. This technique has been used to investigate the adhesion process in early stages of stem cell migration.
Simulation and evaluation of 3D traction force microscopy
Published in Computer Methods in Biomechanics and Biomedical Engineering, 2019
C.N. Holenstein, C.R. Lendi, Nino Wili, J.G. Snedeker
Mechanical interactions of adherent cells with the extracellular matrix (ECM) are a key driver of tissue development and pathogenesis (Iskratsch et al. 2014). It is therefore essential to study not only biomechanical but also mechanical interactions in order to derive an understanding of cellular behavior which involves the acquisition of quantitative information about the forces that are present in this complex microenvironment. Cell-generated forces can be measured with methods commonly referred to as Traction Force Microscopy (TFM) (Dembo et al. 1996), which are most widely applied to cells cultured on flat elastic substrates, and increasingly applied to cells in 3 D culture (Figure 1a).