Electron Microscopy of Surface Antigens on Yeast Protoplasts by the Plasma Polymerization-Film Metal Extraction Replica Method

Author(s):  
A. Tanaka ◽  
M. Yamaguchi ◽  
T. Hirano

The plasma polymerization replica method and its apparatus have been devised by Tanaka (1-3). We have published several reports on its application: surface replicas of biological and inorganic specimens, replicas of freeze-fractured tissues and metal-extraction replicas with immunocytochemical markers.The apparatus for plasma polymerization consists of a high voltage power supply, a vacuum chamber containing a hydrocarbon gas (naphthalene, methane, ethylene), and electrodes of an anode disk and a cathode of the specimen base. The surface replication by plasma polymerization in negative glow phase on the cathode was carried out by gassing at 0.05-0.1 Torr and glow discharging at 1.5-3 kV D.C. Ionized hydrocarbon molecules diffused into complex surface configurations and deposited as a three-dimensionally polymerized film of 1050 nm in thickness.The resulting film on the complex surface had uniform thickness and showed no granular texture. Since the film was chemically inert, resistant to heat and mecanically strong, it could be treated with almost any organic or inorganic solvents.


1997 ◽  
Vol 46 (5) ◽  
pp. 425-430 ◽  
Author(s):  
M. Yamaguchi ◽  
H. Hirokawa ◽  
K. Sugahara ◽  
H. Mizokami ◽  
K. Takeo

Author(s):  
Hirano T. ◽  
M. Yamaguchi ◽  
M. Hayashi ◽  
Y. Sekiguchi ◽  
A. Tanaka

A plasma polymerization film replica method is a new high resolution replica technique devised by Tanaka et al. in 1978. It has been developed for investigation of the three dimensional ultrastructure in biological or nonbiological specimens with the transmission electron microscope. This method is based on direct observation of the single-stage replica film, which was obtained by directly coating on the specimen surface. A plasma polymerization film was deposited by gaseous hydrocarbon monomer in a glow discharge.The present study further developed the freeze fracture method by means of a plasma polymerization film produces a three dimensional replica of chemically untreated cells and provides a clear evidence of fine structure of the yeast plasma membrane, especially the dynamic aspect of the structure of invagination (Figure 1).


1984 ◽  
Vol 30 (8) ◽  
pp. 1008-1013 ◽  
Author(s):  
C. Mouton ◽  
L. Lamonde

Colloidal gold particles 3–6 nm in diameter were prepared and stabilized with the IgG fraction of polyspecific rabbit antisera produced against four different oral bacteria. The immunogold markers were used in homologous reactions to label the bacteria in a preembedding procedure for electron microscopy. An indirect immunofluorescence procedure was concurrently used to optimize the labelling conditions before observation with the electron microscope. The immunogold markers labelled fibrillar structures extending outward 50–275 nm from the Gram-positive cell envelopes and a fuzzy 5–10 nm thick capsulelike layer on the outer aspect of Bacteroides gingivalis. The immunogold method appears to be a simple, rapid, and inexpensive procedure suitable for the study of bacterial surface antigens and can be upgraded with the use of monospecific antibodies.


Nature ◽  
1961 ◽  
Vol 191 (4792) ◽  
pp. 1022-1023 ◽  
Author(s):  
P. F. ELBERS

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