High-resolution scanning electron microscopy for the imaging of nuclear pore complexes and Ran-mediated transport.
High-resolution scanning electron microscopy provides three-dimensional surface images of nuclear pore complexes (NPCs) embedded in the nuclear envelope. Here, we describe a method for exposing the nuclear surface in mammalian tissue culture cells for imaging by scanning electron microscopy. Hypotonic treatment is followed by low-speed centrifugation onto polylysine-coated silicon chips, without the use of detergents. This helps to preserve NPCs close to their native morphology, embedded in undamaged nuclear membranes. This method is particularly advantageous for combining high-resolution imaging of NPCs with mammalian genetic systems.
Faculty of Medicine in the Galilee, Bar-Ilan University, Safed, Israel.
Lihi
L
Shaulov
Boris
B
Fichtman
Amnon
A
Harel
D021581:Active Transport, Cell Nucleus; D000818:Animals; D019556:COS Cells; D002467:Cell Nucleus / Q000378:metabolism* / Q000648:ultrastructure*; D002522:Chlorocebus aethiops; D007038:Hypotonic Solutions / Q000494:pharmacology; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D022022:Nuclear Pore / Q000378:metabolism*; D020931:ran GTP-Binding Protein / Q000378:metabolism*
D007038:Hypotonic Solutions; D020931:ran GTP-Binding Protein