Dual effect of tannic acid on the preservation and ultrastructure of phosphatidyl choline vesicles.
The use of tannic acid has been proposed to improve the preservation of phospholipids in tissues. We investigated the effects of tannic acid on the preservation of small unilamellar vesicles, prepared from sonicated aqueous suspensions of phospholipids. With cryo-electron microscopy it is demonstrated that small unilamellar vesicles are formed after sonication of the phospholipid suspensions. Fixation of vesicles without tannic acid results in extraction of the phospholipids during dehydration and embedding. Fixation of vesicles containing phosphatidyl choline with tannic acid, with or without glutaraldehyde, results in a fast (within a second) aggregation of the vesicles and the resulting sediment can be dehydrated and embedded when a postfixation in osmium tetroxide is carried out. Small unilamellar vesicles fixed in this way are retrieved in thin sections as multilamellar vesicles with a periodicity of about 5 nm for dimyristoylphosphatidyl choline and about 6 nm for dioleoylphosphatidyl choline. By using 14C-phosphatidyl choline it was also demonstrated that tannic acid prevents to a large extent the extraction of phosphatidyl choline during fixation, dehydration and embedding. This dual effect of tannic acid on phosphatidyl choline, aggregation and fixation, should be considered when using tannic acid in tissue preparation.
Schrijvers
A H
AH
Department of Physiology, University of Limburg, Maastricht, The Netherlands.
Frederik
P M
PM
Stuart
M C
MC
van der Vusse
G J
GJ
Reneman
R S
RS
D005615:Freezing; D005976:Glutaral; D047348:Hydrolyzable Tannins; D008567:Membranes, Artificial; D008854:Microscopy, Electron; D009993:Osmium Tetroxide; D010713:Phosphatidylcholines; D011309:Preservation, Biological; D013634:Tannins
D047348:Hydrolyzable Tannins; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D013634:Tannins; D009993:Osmium Tetroxide; D005976:Glutaral