@article{878d51438c0540698e7f8b39a5c5d969,
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.",
author = "Welf, {Erik S.} and Driscoll, {Meghan K.} and Dean, {Kevin M.} and Claudia Sch{\"a}fer and Jun Chu and Davidson, {Michael W.} and Lin, {Michael Z.} and Gaudenz Danuser and Reto Fiolka",
note = "Funding Information: We would like to thank John Hammer for the GFP-tractin construct, Magnus H{\"o}{\"o}k for the CNA35 peptide, John Minna for the Kras V12 construct, Bob Fischer for the td-Tomato membrane marker construct, Wesley Burford and Kim Reed for molecular biology support, and Sean Morrison, Elena Piskounova, and Ugur Eskiocak for helping to establish the workflow for culturing primary melanoma. Some of the core functions and routines in the microscope control software are licensed under an MTA from Howard Hughes Medical Institute, Janelia Farm Research Campus. The full microscope control software code can be requested from the corresponding authors and will be distributed under MTAs with HHMI and UT Southwestern Medical Center. E.S.W., M.K.D., K.M.D., C.S., R.F., and G.D. were supported by Cancer Prevention Research Institute of Texas grant R1225 to G.D., and M.Z.L. and J.C. were supported by NIH grant 5DP1GM111003-02 . M.K.D. was also supported by NIH grant F32-GM116370 . Funding Information: We would like to thank John Hammer for the GFP-tractin construct, Magnus H{\"o}{\"o}k for the CNA35 peptide, John Minna for the KrasV12 construct, Bob Fischer for the td-Tomato membrane marker construct, Wesley Burford and Kim Reed for molecular biology support, and Sean Morrison, Elena Piskounova, and Ugur Eskiocak for helping to establish the workflow for culturing primary melanoma. Some of the core functions and routines in the microscope control software are licensed under an MTA from Howard Hughes Medical Institute, Janelia Farm Research Campus. The full microscope control software code can be requested from the corresponding authors and will be distributed under MTAs with HHMI and UT Southwestern Medical Center. E.S.W., M.K.D., K.M.D., C.S., R.F., and G.D. were supported by Cancer Prevention Research Institute of Texas grant R1225 to G.D., and M.Z.L. and J.C. were supported by NIH grant 5DP1GM111003-02. M.K.D. was also supported by NIH grant F32-GM116370. Publisher Copyright: {\textcopyright} 2016 Elsevier Inc.",
year = "2016",
month = feb,
day = "22",
doi = "10.1016/j.devcel.2016.01.022",
language = "English (US)",
volume = "36",
pages = "462--475",
journal = "Developmental cell",
issn = "1534-5807",
publisher = "Cell Press",
number = "4",
}