Electron microscopy of biological specimens by the plasma-polymerization rapid-freeze replica method

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


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).


2017 ◽  
Vol 2017 ◽  
pp. 1-10 ◽  
Author(s):  
Carlos Andrés Covarrubias-Gordillo ◽  
Florentino Soriano-Corral ◽  
Carlos Alberto Ávila-Orta ◽  
Victor Javier Cruz-Delgado ◽  
María Guadalupe Neira-Velázquez ◽  
...  

Carbon nanofibers (CNFs), graphene platelets (GPs), and their mixtures were treated by plasma polymerization of propylene. The carbon nanoparticles (CNPs) were previously sonicated in order to deagglomerate and increase the surface area. Untreated and plasma treated CNPs were analyzed by dynamic light scattering (DLS), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, and thermogravimetric analysis (TGA). DLS analysis showed a significant reduction of average particle size, due to the sonication pretreatment. Plasma polymerized propylene was deposited on the CNPs surface; the total amount of polymerized propylene was from 4.68 to 6.58 wt-%. Raman spectroscopy indicates an increase in the sp3 hybridization of the treated samples, which suggest that the polymerized propylene is grafted onto the CNPs.


2010 ◽  
Vol 9 ◽  
pp. 45-53 ◽  
Author(s):  
E. Hernández-Hernández ◽  
M.G. Neira-Velázquez ◽  
L.F. Ramos de Valle ◽  
Arturo Ponce ◽  
D. Weinkauf

Carbon nanofibers (CNF) were surface coated with a thin layer of poly (acrylic acid) (PAA) via plasma polymerization of acrylic acid (AA). The plasma polymerization treated CNF resulted in a significant change of its surface characteristics from hydrophobic to hydrophilic due to the polar groups in PAA-coating layer. Dispersion tests in water and ethanol confirmed a marked change in the hydrophobicity of the treated CNF. In addition, FTIR results showed a new signal for the treated CNF at 1046 cm-1, assigned to C-O groups and represented the presence of PAA. Morphology results by scanning electron microscopy (SEM) revealed that the untreated CNF has a very smooth surface, whereas the treated CNF showed certain roughness on its surface due to the deposited PAA coating layer.PAA coating on the treated CNF was also observed by transmission electron microscopy (TEM) and its thickness was determined to be between 14 to 18 nm. Dynamic Light Scattering (DLS) was used to show that plasma polymerization improved the dispersion of treated CNF and prevented its agglomeration in water versus the poor dispersion observed with untreated CNF.


Author(s):  
Sumire Inaga ◽  
Hitoshi Osatake ◽  
Akihiro lino ◽  
Keiichi Tanaka

So far, the ultrastructure of DNA strand and nucleosome had been observed mainly by transmission electron microscopy with some techniques (thin-sectioning, spreading method, replica method and so on). Among them, the freeze-etching replica method gave high magnified images of DNA double helix (Ruben et al., 1989). Further, scanning tunneling microscopy also elucidated the images of major and minor grooves in a helical DNA duplex. Though scanning electron microscopy (SEM) was also applied for observing chromatin structures, it had been difficult to observe clearly such small materials. Because, the resolution of SEM was too poor to investigate such fine structures. The obstruction of resolution, however, was overcome by the development of an ultrahigh resolution SEM (UHS-T1, Tanaka et al., 1985). Using the SEM, we could successfully observed naked DNA strands and nucleosomes of chicken erythrocyte nuclei without any metal-coating.Preparations were made by the microspreading procedure basically according to the method of Seki et al.


Sign in / Sign up

Export Citation Format

Share Document