High-resolution molecular localization by freeze-fracture replica labeling.
The freeze-fracture technique splits the frozen lipid bilayer membrane into two halves and immobilizes membrane proteins and lipids by the vacuum evaporation of platinum and carbon. After a treatment by SDS to remove extramembrane materials, the specimen is subjected to immunogold labeling, which gives information on the two-dimensional distribution of membrane molecules and their relationship to various differentiated structures. In combination with rapid freezing, the freeze-fracture technique has an advantage over other methods using conventional chemical fixation because the distribution of lipids as well as proteins can be observed at the mesoscale in a wide area of the membrane.
Fujita
Akikazu
A
Department of Anatomy and Molecular Cell Biology, Nagoya University Graduate School of Medicine, Nagoya, Japan.
Fujimoto
Toyoshi
T
D002244:Carbon / Q000737:chemistry; D002478:Cells, Cultured; D005614:Freeze Fracturing / Q000379:methods*; D017979:Gold Colloid / Q000737:chemistry / Q000276:immunology; D006801:Humans; D008051:Lipid Bilayers / Q000737:chemistry; D008563:Membrane Lipids / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry*; D010984:Platinum / Q000737:chemistry; D016628:Replica Techniques / Q000379:methods*; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods*
D017979:Gold Colloid; D008051:Lipid Bilayers; D008563:Membrane Lipids; D008565:Membrane Proteins; D012967:Sodium Dodecyl Sulfate; D010984:Platinum; D002244:Carbon