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Dev Cell. 2016 Feb 22;36(4):462-75. doi: 10.1016/j.devcel.2016.01.022.

Quantitative Multiscale Cell Imaging in Controlled 3D Microenvironments.

Author information

1
Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
2
Departments of Bioengineering and Pediatrics, Stanford University, Stanford, CA 94305, USA.
3
National High Magnetic Field Laboratory, Department of Biological Science, Florida State University, Tallahassee, FL 32310, USA.
4
Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA. Electronic address: gaudenz.danuser@utsouthwestern.edu.
5
Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA. Electronic address: reto.fiolka@utsouthwestern.edu.

Abstract

The microenvironment determines cell behavior, but the underlying molecular mechanisms are poorly understood because quantitative studies of cell signaling and behavior have been challenging due to insufficient spatial and/or temporal resolution and limitations on microenvironmental control. Here we introduce microenvironmental selective plane illumination microscopy (meSPIM) for imaging and quantification of intracellular signaling and submicrometer cellular structures as well as large-scale cell morphological and environmental features. We demonstrate the utility of this approach by showing that the mechanical properties of the microenvironment regulate the transition of melanoma cells from actin-driven protrusion to blebbing, and we present tools to quantify how cells manipulate individual collagen fibers. We leverage the nearly isotropic resolution of meSPIM to quantify the local concentration of actin and phosphatidylinositol 3-kinase signaling on the surfaces of cells deep within 3D collagen matrices and track the many small membrane protrusions that appear in these more physiologically relevant environments.

PMID:
26906741
PMCID:
PMC4784259
DOI:
10.1016/j.devcel.2016.01.022
[Indexed for MEDLINE]
Free PMC Article

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