A reliable method for obtaining matched replicas of freeze-fractured cell suspensions. 1980

M H Wilkinson, and D H Northcote

A method that overcomes the majority of practical problems involved in performing matched freeze-fracture replication of cell suspensions is described. Specimens are rapidly frozen between copper support plates. The prior attachment of gold grids to the plates using Formvar ensures the exact alignment of corresponding grid squares and the production of a central fissure running through the specimen upon fracturing. Grids, with replicas permanently attached, are removed from the support plates by dissolving the Formvar. Sample digestion and rinsing of replicas is by a gentle procedure employing glass capillaries as a means of handling the grids. Matched replication of plant and animal cells has been achieved by this methods. Apart from the excellent preservation of the membrane ultrastructure of unfixed cells and the capacity for matching both membrane halves this technique also enhances the fracture characteristics of certain specimens, notably isolated protoplasts, providing larger fracture faces than are obtained by the knife-splintering method.

UI MeSH Term Description Entries
D010944 Plants Multicellular, eukaryotic life forms of kingdom Plantae. Plants acquired chloroplasts by direct endosymbiosis of CYANOBACTERIA. They are characterized by a mainly photosynthetic mode of nutrition; essentially unlimited growth at localized regions of cell divisions (MERISTEMS); cellulose within cells providing rigidity; the absence of organs of locomotion; absence of nervous and sensory systems; and an alternation of haploid and diploid generations. It is a non-taxonomical term most often referring to LAND PLANTS. In broad sense it includes RHODOPHYTA and GLAUCOPHYTA along with VIRIDIPLANTAE. Plant
D011523 Protoplasts The protoplasm and plasma membrane of plant, fungal, bacterial or archaeon cells without the CELL WALL. Protoplast
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
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

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