Quantitative picture analysis of freeze-fracture electron-micrographs. 1981

H Gruler

A method of three-dimensional reconstruction of the surface profile of artificial and natural membranes from freeze quenched electron micrographs is presented. The direct relation between the Pt-layer thickness and the local orientation of the membrane allows a reconstruction of the surface. The efficiency of this method is demonstrated on the quantitative analysis of some fine structures. These essential results are: 1. In the low resolution observation structural elements of a yeast cell were quantitatively described, (i) The diameter of a yeast cell is determined (4.2 microgram). (ii) The cell wall thickness is measured (150 nm). (iii) The dimension of cell wall incapsulated vesicles is determined (60-80 nm). (iv) The damlike protrusion in the plasma membrane has a triangular cross section. The height is 23 nm and the half width 50 nm. The particle assembly in the damlike protrusion is in a crystalline state. The change in surface curvature is probable due to a phase separation of a biaxial cluster in an uniaxial membrane. (v) Membrane bound particles can be distinguished by their surface profiles. 2. The resolution of surface profiles is limited by the size of the platinum grain. An average procedure can lead to a resolution of 0.2 nm. This increase is resolution can be understood with the uncertainty relation: The uncertainty of the profile in one dimension times the uncertainty in the other dimension (averaging length) is the area of the platinum grain. The monolayer thickness of dipalmitoyl phosphatidyl choline and dimyristoyl choline are distinguishable 2.6 +/- 0.2 nm and 2.4 +/- 0.2 nm respectively. The surface profile of a two-dimensional crystal in the membrane of a yeast cell can be determined with high accuracy. The two profiles of the inner and outer monolayer do not fit exactly together. A part of the membrane bound particle is pulled out of the monolayer during the fracturing procedure. 3. The third part investigates special fluctuation of the surface. (i) The mixture of dipalmitoyl phosphatidyl choline and dioleyl phosphatidyl choline shows a periodic structure. The fluctuation besides this periodicity can be explained by a spinodal decomposition during cryofixation. (ii) The fluctuation of a periodic structure can also be induced by thermal motion. The fluctuation of dimyristol phosphatidyl choline quenched from a temperature between the pre- and maintransition determines only one kind of elastic constant. This curvature elastic constant is in the order of 10(-20) Joule. (iii) The fluctuation of the particle density can be related with the particle-particle compressibility. We choose the clusters induced by polylysine in a membrane with charged and uncharged lipids as particles. The compressibility is in the order of 10(-6) Newton/m which is comparable to those of a monolayer in a gaseous state.

UI MeSH Term Description Entries
D008081 Liposomes Artificial, single or multilaminar vesicles (made from lecithins or other lipids) that are used for the delivery of a variety of biological molecules or molecular complexes to cells, for example, drug delivery and gene transfer. They are also used to study membranes and membrane proteins. Niosomes,Transferosomes,Ultradeformable Liposomes,Liposomes, Ultra-deformable,Liposome,Liposome, Ultra-deformable,Liposome, Ultradeformable,Liposomes, Ultra deformable,Liposomes, Ultradeformable,Niosome,Transferosome,Ultra-deformable Liposome,Ultra-deformable Liposomes,Ultradeformable Liposome
D008433 Mathematics The deductive study of shape, quantity, and dependence. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed) Mathematic
D008854 Microscopy, Electron Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen. Electron Microscopy
D010743 Phospholipids Lipids containing one or more phosphate groups, particularly those derived from either glycerol (phosphoglycerides see GLYCEROPHOSPHOLIPIDS) or sphingosine (SPHINGOLIPIDS). They are polar lipids that are of great importance for the structure and function of cell membranes and are the most abundant of membrane lipids, although not stored in large amounts in the system. Phosphatides,Phospholipid
D002462 Cell Membrane The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells. Plasma Membrane,Cytoplasmic Membrane,Cell Membranes,Cytoplasmic Membranes,Membrane, Cell,Membrane, Cytoplasmic,Membrane, Plasma,Membranes, Cell,Membranes, Cytoplasmic,Membranes, Plasma,Plasma Membranes
D002784 Cholesterol The principal sterol of all higher animals, distributed in body tissues, especially the brain and spinal cord, and in animal fats and oils. Epicholesterol
D005614 Freeze Fracturing Preparation for electron microscopy of minute replicas of exposed surfaces of the cell which have been ruptured in the frozen state. The specimen is frozen, then cleaved under high vacuum at the same temperature. The exposed surface is shadowed with carbon and platinum and coated with carbon to obtain a carbon replica. Fracturing, Freeze,Fracturings, Freeze,Freeze Fracturings
D012441 Saccharomyces cerevisiae A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement. Baker's Yeast,Brewer's Yeast,Candida robusta,S. cerevisiae,Saccharomyces capensis,Saccharomyces italicus,Saccharomyces oviformis,Saccharomyces uvarum var. melibiosus,Yeast, Baker's,Yeast, Brewer's,Baker Yeast,S cerevisiae,Baker's Yeasts,Yeast, Baker
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