Improved cryofixation applicable to freeze etching. 1971

L Bachmann, and W W Schmitt

Freeze etching of solute model systems (e.g., glycerol or ferritin solutions) demonstrates that cryofixation can introduce serious artifacts due to the segregation of the dissolved or dispersed material from the solvent. Since, in principle, this problem can be reduced by increasing the cooling rate, a new technique has been developed which combines spray freezing with freeze etching. This spray-freeze-etching is applied by first spraying the specimen into a liquid cryomedium. The frozen droplets are then "glued" together with butylbenzene to form a regular freeze-etch specimen, while the temperature of the sample is kept at -85 degrees C. The results obtained by spray-freeze-etching are far superior to those obtained by standard freezing. Our results, using 5% glycerol as a test specimen, are equivalent to those obtained by the high-pressure method (1). The reduction of segregation during freezing makes freeze etching a method applicable for the investigation of solute systems. Furthermore, the study of unicellular organisms or cellular fractions by freeze etching without the use of antifreeze is made possible.

UI MeSH Term Description Entries
D008722 Methods A series of steps taken in order to conduct research. Techniques,Methodological Studies,Methodological Study,Procedures,Studies, Methodological,Study, Methodological,Method,Procedure,Technique
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
D005613 Freeze Etching A replica technique in which cells are frozen to a very low temperature and cracked with a knife blade to expose the interior surfaces of the cells or cell membranes. The cracked cell surfaces are then freeze-dried to expose their constituents. The surfaces are now ready for shadowing to be viewed using an electron microscope. This method differs from freeze-fracturing in that no cryoprotectant is used and, thus, allows for the sublimation of water during the freeze-drying process to etch the surfaces. Etching, Freeze
D005615 Freezing Liquids transforming into solids by the removal of heat. Melting
D006652 Histological Techniques Methods of preparing tissue for examination and study of the origin, structure, function, or pathology. Histologic Technic,Histologic Technics,Histologic Technique,Histologic Techniques,Histological Technics,Technic, Histologic,Technics, Histologic,Technique, Histologic,Techniques, Histologic,Histological Technic,Histological Technique,Technic, Histological,Technics, Histological,Technique, Histological,Techniques, Histological

Related Publications

L Bachmann, and W W Schmitt
January 1969, Histochemie. Histochemistry. Histochimie,
L Bachmann, and W W Schmitt
April 1972, Laboratory practice,
L Bachmann, and W W Schmitt
January 1969, International review of cytology,
L Bachmann, and W W Schmitt
January 2014, Methods in molecular biology (Clifton, N.J.),
L Bachmann, and W W Schmitt
October 1975, Science (New York, N.Y.),
L Bachmann, and W W Schmitt
January 1969, Cryobiology,
L Bachmann, and W W Schmitt
January 1993, Methods in cell biology,
Copied contents to your clipboard!