#Transformice map database how to

Various experimental techniques for quantitative traction force mapping at spatial scales ranging from multicellular sheets to single molecules have been developed over the last 30 years. Therefore, the ability to measure cellular traction forces is critical to better understand the cellular and molecular mechanisms behind many basic biological processes at both the cell and tissue levels. Cellular traction forces are also critical for pathological processes, such as cancer metastasis ( Wirtz, Konstantopoulos, & Searson, 2011). These forces allow cells to perform vital physiological tasks during embryo morphogenesis, wound healing, and the immune response ( DuFort, Paszek, & Weaver, 2011). A brief discussion of different ways to visualize and analyze the results serves to illustrate possible uses of high-resolution TFM in biomedical research.Ĭell contractile forces generated by the actomyosin cytoskeleton and transmitted to the extracellular matrix (ECM) through integrin-based focal adhesions drive cell adhesion, spreading, and migration.

#Transformice map database how to

We demonstrate the application of the algorithm and explain how to choose a regularization parameter for suppression of the measurement error. We describe the implementation of a new algorithm for processing of images of fiducial markers that are taken below the surface of the substrate, which significantly improves data quality. In addition, we provide the theoretical background of traction reconstruction and experimental considerations important to design a high-resolution TFM experiment. We describe fabrication of polyacrylamide substrates labeled with multiple colors of fiducial markers, functionalization of the substrates with ECM proteins, setting up the experiment, and imaging procedures.

Here, we provide detailed protocols for measuring cellular forces exerted on two-dimensional elastic substrates with a high-resolution traction force microscopy (TFM) method.

However, quantitative mapping of these forces has been difficult since there has been no experimental technique to visualize nanonewton forces at submicrometer spatial resolution.

Cellular forces generated by the actomyosin cytoskeleton and transmitted to the extracellular matrix (ECM) through discrete, integrin-based protein assemblies, that is, focal adhesions, are critical to developmental morphogenesis and tissue homeostasis, as well as disease progression in cancer.