Journal Article

Technical Advance: An Automated Device for Cryofixation of Specimens of Electron Microscopy using Liquid Helium

Akiko Hisada, Tomoko Yoshida, Shigeo Kubota, Naoko K. Nishizawa and Masaki Furuya

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 42, issue 9, pages 885-893
Published in print September 2001 | ISSN: 0032-0781
Published online September 2001 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pce132
Technical Advance: An Automated Device for Cryofixation of
Specimens of Electron Microscopy using Liquid
Helium

More Like This

Show all results sharing these subjects:

  • Biochemistry
  • Molecular and Cell Biology
  • Plant Sciences and Forestry

GO

Show Summary Details

Preview

Metal-contact rapid freezing using liquid helium is theoretically the best method for preserving the fine structure of living cells with high temporal resolution in preparation of tissue samples for electron microscopy. However, this method is not commonly used, because of its technical difficulty and low reproducibility. We have designed and constructed an automatic device which allows simple, rapid and reproducible preparation of high-quality electron microscopic specimens by the non-specialist. We assessed the quality of cryofixation in samples prepared using this device by examining the preservation of cellular ultrastructure in relation to distance from the freezing block, and found that the region within 10 µm of the metal-contact plane was fixed with the highest quality. We applied this device, in combination with freeze-substitution methods and immunocytochemical techniques, to two phenomena involving rapid movement of subcellular components: (1) active movement of subcellular structures in the papillar cells of stigma and (2) light-induced rapid subcellular translocation of phytochrome A. Considering the importance of understanding subcellular processes of living cells for molecular and cell biology, this device will be a useful tool for diverse biological applications in the near future.

Keywords: Key words: Arabidopsis — Automated metal-contact cryofixation — Electron microscopy — Immunostaining — Liquid helium — Pea.; Abbreviations: BSA, bovine serum albumin; GFP, green fluorescent protein; PBS, phosphate-buffered saline; PHYA, apoprotein of phytochrome A; TEM, transmission electron microscope.

Journal Article.  5393 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

Full text: subscription required

How to subscribe Recommend to my Librarian

Users without a subscription are not able to see the full content. Please, subscribe or login to access all content.