Extraction of lipids during freeze-substitution of Acholeplasma laidlawii-cells for electron microscopy.
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.
D000128:Acholeplasma laidlawii / Q000648:ultrastructure*; D005615:Freezing; D005976:Glutaral; D007202:Indicators and Reagents; D008055:Lipids / Q000302:isolation & purification*; D008854:Microscopy, Electron / Q000379:methods
D007202:Indicators and Reagents; D008055:Lipids; D005976:Glutaral