title: Quantitative Multiscale Cell Imaging in Controlled 3D Microenvironments.
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.
authors.affiliation: Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
authors.forename: Erik S
authors.initials: ES
authors.lastname: Welf
authors.affiliation: Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
authors.forename: Meghan K
authors.initials: MK
authors.lastname: Driscoll
authors.affiliation: Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Dean
authors.affiliation: Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
authors.forename: Claudia
authors.initials: C
authors.lastname: Schäfer
authors.affiliation: Departments of Bioengineering and Pediatrics, Stanford University, Stanford, CA 94305, USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Chu
authors.affiliation: National High Magnetic Field Laboratory, Department of Biological Science, Florida State University, Tallahassee, FL 32310, USA.
authors.forename: Michael W
authors.initials: MW
authors.lastname: Davidson
authors.affiliation: Departments of Bioengineering and Pediatrics, Stanford University, Stanford, CA 94305, USA.
authors.forename: Michael Z
authors.initials: MZ
authors.lastname: Lin
authors.affiliation: Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA. Electronic address: gaudenz.danuser@utsouthwestern.edu.
authors.forename: Gaudenz
authors.initials: G
authors.lastname: Danuser
authors.affiliation: Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA. Electronic address: reto.fiolka@utsouthwestern.edu.
authors.forename: Reto
authors.initials: R
authors.lastname: Fiolka
mesh_terms: D000199:Actins / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002465:Cell Movement / Q000502:physiology*; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D006801:Humans; D008853:Microscopy / Q000379:methods; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*
chemical_list: D000199:Actins; D003094:Collagen; D019869:Phosphatidylinositol 3-Kinases