Exploring membrane domains using native membrane sheets and transmission electron microscopy.
The flow of information in cells requires the constant remodeling of cell signaling and trafficking networks. To observe the remodeling events associated with activation of receptors on the cell surface, the authors have generated and analyzed high-resolution topographical maps of colloidal gold nanoprobes (3-10 nm) marking receptors, signaling proteins, and lipids in native membranes. The technology involves sandwiching of cells between glass cover slips and electron microscopy (EM) grids, followed by ripping. Membrane sheets on EM grids are fixed, labeled with functionalized nanoprobes, and imaged by transmission electron microscopy. Probe coordinates are extracted from digitized images and the distributions of the probes are analyzed with respect to each other and to membrane features like clathrin-coated pits, caveolae, and the cortical cytoskeleton.
Wilson
Bridget S
BS
Dept. of Pathology and Cancer Center, University of New Mexico School of Medicine, Albuquerque, USA.
Pfeiffer
Janet R
JR
Raymond-Stintz
Mary Ann
MA
Lidke
Diane
D
Andrews
Nicholas
N
Zhang
Jun
J
Yin
Wenxia
W
Steinberg
Stanly
S
Oliver
Janet M
JM
D000465:Algorithms; D021941:Caveolae / Q000648:ultrastructure; D002460:Cell Line; D045744:Cell Line, Tumor; D002966:Clathrin / Q000648:ultrastructure; D022163:Clathrin-Coated Vesicles / Q000648:ultrastructure; D016000:Cluster Analysis; D003599:Cytoskeleton / Q000648:ultrastructure; D005898:Glass; D006801:Humans; D021962:Membrane Microdomains / Q000737:chemistry* / Q000648:ultrastructure*; D046529:Microscopy, Electron, Transmission / Q000295:instrumentation / Q000379:methods*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000648:ultrastructure; D017455:Receptors, IgE / Q000648:ultrastructure; D013269:Stochastic Processes
D002966:Clathrin; D018719:Erb-b2 Receptor Tyrosine Kinases; D017455:Receptors, IgE