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

• Published in: Journal of electron microscopy Vol. 46; no. 5; pp. 425 - 430
• Main Authors: Yamaguchi, Masashi, Hirokawa, Hideo, Sugahara, Keishin, Mizokami, Hiroshi, Takeo, Kanji
• Format: Journal Article
• Language: English
• Published: Japan Oxford University Press 1997
• Subjects: Aluminum Silicates; Cryopreservation - methods; Crystallization; Hepatitis B virus - ultrastructure; hepatitis B virus core particle; Magnesium Oxide; mica flakes method; Microscopy, Electron - methods; Microtomy; Negative Staining; Plasma; plasma polymerization; Polymers; rapid freeze; replica; Replica Techniques; Specimen Handling - methods; T-Phages - ultrastructure; T4 bacteriophage
• DOI: 10.1093/oxfordjournals.jmicro.a023538
• EISSN: 1477-9986
• ISSN: 0022-0744

The plasma-polymerization replica method is a unique replica technique for transmission electron microscopy. In the present study, we used this method in combination with a rapid-freeze technique to observe T4 bacteriophages and hepatitis B virus core particles. The heads of T4 bacteriophages appeared hexagonal and measured –110 nm in length. Striations in their tails were also visible, indicating that the resolution of the present method is better than 4 nm. The images corresponded well with those obtained by ice-embedding and negative staining methods, with respect to both morphology and size of the phage particle. Hepatitis B virus core particles observed by the present method appeared round, –30 nm in diameter, with hollow centres. Again, the morphology and size of the particles corresponded well with those obtained by ice-embedding, negative staining, and ultrathin sectioning. From these results, we conclude that the plasma-polymerization rapid-freeze replica method provides a useful technique for observation of biological specimens in a natural state and at high resolution.