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Extraction of lipids during freeze‐substitution of Acholeplasma laidlawii ‐cells for electron microscopy
Author(s) -
Weibull C.,
Villiger W.,
Carlemalm E.
Publication year - 1984
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.1984.tb02513.x
Subject(s) - acetone , methanol , electron microscope , extraction (chemistry) , liquid nitrogen , chemistry , bacteria , chromatography , biochemistry , organic chemistry , biology , physics , optics , genetics
SUMMARY Cells of the bacterium Acholeplasma laidlawii were rapidly frozen against a copper block cooled by liquid helium. The frozen cells were transferred to liquid nitrogen and subsequently to acetone or methanol at 183 K. After 24 h the cells were infiltrated at 203 K with a non‐polar methacrylate resin of the same type as Lowicryl HM20. The resin was cured at the same temperature. Acetone extracted approximately 5% of the lipid content of the cells, methanol 15–45% and the resin only negligible amounts. Similar results were obtained with A. laidlawii ‐ghosts. The cells appeared well preserved when examined in the electron microscope.