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Journal of Lipid Research, Vol. 9, 416-424, July 1968
Copyright © 1968 by Lipid Research, Inc.

Lipid fixation during preparation of chloroplasts for electron microscopy

Alpaslan Ongun , W. W. Thomson , and J. B. Mudd

Department of Biochemistry, Department of Life Sciences, and Statewide Air Pollution Research Center, University of California, Riverside, California 92502

Reaction of osmium tetroxide with isolated spinach chloroplasts fixed completely the glycolipids, phosphatidyl glycerol, and phosphatidyl choline. Under the same reaction conditions only 30% of the chlorophyll was fixed.

Reaction of potassium permanganate with isolated spinach chloroplasts fixed more than 90% of the glycolipids, phosphatidyl glycerol, and phosphatidyl choline, provided the reaction period was long enough. Potassium permanganate also fixed the chlorophyll.

Reaction of osmium tetroxide and potassium permanganate with isolated 14C-lipids from Chlorella pyrenoidosa fixed 59% and 66% of the radioactivity, respectively. The lipids that were not fixed included sterols and pigments.

Electron micrographs show that chloroplasts extracted with chloroform-methanol after fixation in osmium tetroxide or potassium permanganate differ from those dehydrated with acetone mainly in that in the former, osmiophilic globules have been removed and there seems to be some fusion of the boundary membranes and grana membranes. These effects may be due to the extraction of unfixed, neutral lipids such as sterols and quinones.

Supplementary key words chloroplasts • lipids • extraction • fixation • osmium tetroxide • potassium permanganate • electron microscopy • osmiophilic globules • membranes

Submitted on December 18, 1967
Accepted on March 5, 1968


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