A Quick Preparative Method for Electron Microscopy Observations of Delicate Objects Using Alginate Embedding Medium |
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| Authors: | Christian Tamponnet Jean-Noë l Barbotin Fran oise Piton |
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| Affiliation: | a Laboratory of Membrane Biology, University of Paris VII, Paris, Cedex, Franceb Laboratory of Enzyme Technology, UA CNRS 523, Compiegne University of Technology, Compiegne, Cedex, Francec Department of Biological Sciences, Life Sciences Building, Room 372, State University of New York at Stony Brook, Stony Brook, NY |
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| Abstract: | A quick, safe method has been devised for embedding small or fragile specimens and keeping delicate structures intact. Cells or organisms to be embedded are placed in a viscous sodium alginate solution (1-2%), which is then polymerized in 100 mM calcium chloride. The resulting gel is easily dehydrated, embedded in resin and sectioned for electron microscopy. This method, the alginate gel portion of which was originally developed for the immobilization of Euglena, allows direct observation of each element of the specimens in micrographs. If desired, the alginate can be removed after sectioning by sequestration of calcium in a 20 mM solution of sodium citrate or a 10 mM solution of EGTA. Cells and organelles in the sections respond normally to standard staining procedures. |
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