DOI: 10.1098/rstb.2008.2268. Phase contrast electron microscopy: development of thin-film phase plates and biological applications

Phase contrast electron microscopy: development of thin-film phase plates and biological applications

10.1098/rstb.2008.2268

Crossref journal-article

The Royal Society

Philosophical Transactions of the Royal Society B: Biological Sciences (175)

Abstract

Phase contrast transmission electron microscopy (TEM) based on thin-film phase plates has been developed and applied to biological systems. Currently, development is focused on two techniques that employ two different types of phase plates. The first technique uses a Zernike phase plate, which is made of a uniform amorphous carbon film that completely covers the aperture of an objective lens and can retard the phase of electron waves byπ/2, except at the centre where a tiny hole is drilled. The other technique uses a Hilbert phase plate, which is made of an amorphous carbon film that is twice as thick as the Zernike phase plate, covers only half of the aperture and retards the electron wave phase byπ. By combining the power of efficient phase contrast detection with the accurate preservation achieved by a cryotechnique such as vitrification, macromolecular complexes and supermolecular structures inside intact bacterial or eukaryotic cells may be visualized without staining. Phase contrast cryo-TEM has the potential to bridge the gap between cellular and molecular biology in terms of high-resolution visualization. Examples using proteins, viruses, cyanobacteria and somatic cells are provided.

Bibliography

Nagayama, K., & Danev, R. (2008). Phase contrast electron microscopy: development of thin-film phase plates and biological applications. Philosophical Transactions of the Royal Society B: Biological Sciences, 363(1500), 2153â2162.

Authors 2

Kuniaki Nagayama (first)
Radostin Danev (additional)

References 45 Referenced 59

10.1515/zna-1970-0532 / Z. Naturforschg / Der Einfluβ einer streuenden Phasenplatte auf das elektronen mikroskopische Bild by Badde H.G (1970)
10.1073/pnas.230282097
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Born M& Wolf E Principle of optics. 7th edn. 1999 Cambridge UK:Cambridge University Press.
10.1016/j.jsb.2005.08.004
10.1143/JPSJ.70.696
10.1016/S0304-3991(01)00088-2
10.1143/JPSJ.73.2718
10.2142/biophysics.2.35
10.1016/j.jsb.2007.10.015
10.1023/A:1021234621466
10.1111/j.1365-2818.1982.tb04625.x
Faget J. Fagot M. Ferre J. & Fert C. 1962 Microscopie electronique a contraste de phase. In Proc. 5th Int. Cong. Electron Microscopy A-7 . London UK: Academic Press.
10.1111/j.1749-6632.1960.tb49990.x
10.1016/0304-3991(91)90159-4
10.1016/0304-3991(82)90194-2
10.1016/S0022-2836(83)80179-X
10.1083/jcb.81.2.275
{'key': 'e_1_3_2_20_1', 'first-page': '317', 'article-title': 'Transfer doublet and an elaborated phase plate holder for 120\u200akV electron-phase microscope', 'volume': '54', 'author': 'Hosokawa F', 'year': '2005', 'journal-title': 'J. Electron Microsc'} / J. Electron Microsc / Transfer doublet and an elaborated phase plate holder for 120 kV electron-phase microscope by Hosokawa F (2005)
10.1016/j.jsb.2005.12.004
10.1111/j.1365-2818.1973.tb03799.x
10.1128/JB.188.2.805-808.2006
{'key': 'e_1_3_2_24_1', 'first-page': '79', 'article-title': 'Observation of in vivo DNA in ice embedded whole cyanobacterial cells by Hilbert differential contrast transmission electron microscopy (HDC-TEM)', 'volume': '54', 'author': 'Kaneko Y', 'year': '2007', 'journal-title': 'Plasma Fusion Res'} / Plasma Fusion Res / Observation of in vivo DNA in ice embedded whole cyanobacterial cells by Hilbert differential contrast transmission electron microscopy (HDC-TEM) by Kaneko Y (2007)
10.1093/jmicro/dfh105
{'key': 'e_1_3_2_26_1', 'first-page': '245', 'article-title': 'Phase contrast in electron microscope images with an electrostatic phase plate', 'volume': '44', 'author': 'Krakow W', 'year': '1975', 'journal-title': 'Optik'} / Optik / Phase contrast in electron microscope images with an electrostatic phase plate by Krakow W (1975)
10.1063/1.1755023
10.1016/j.str.2004.05.006
10.1016/j.jmb.2006.12.043
10.1143/JPSJ.68.811
10.1016/S1076-5670(05)38002-5
Nomarski G. 1952 Interferométre á polarization French Patent 1. 059. 123.
10.1364/AO.11.002840
10.1016/S0092-8674(01)00617-1
Reimer L Transmission electron microscopy. 4th edn. 1997 Berlin Germany:Springer. (10.1007/978-3-662-14824-2)
10.1038/nsmb1046
10.1063/1.1698233
10.1007/s00795-006-0341-8
Smith F. H. 1947 Microscopes British Patent 639 014 Class 97 (i) CroupXX.
Sugawara K. Danev R. Yamaguchi M. Nishiyama K. & Nagayama K. 2005 Phase contrast transmission the electron microscopy of rapidly frozen ice-embedded influenza A virus. Cytologia 70 No. 4 cover description.
10.1016/S0022-5320(76)80099-8
10.1002/bbpc.19700741110 / Bunsen-Gesellschaft / An electrostatic phase plate for the electron microsope by Unwin P.N.T (1970)
10.1002/ar.1091490307
{'key': 'e_1_3_2_44_1', 'first-page': '17', 'article-title': 'High resolution electron microscopy with profiled phase plates', 'volume': '44', 'author': 'Willasch D', 'year': '1975', 'journal-title': 'Optik'} / Optik / High resolution electron microscopy with profiled phase plates by Willasch D (1975)
Yamaguchi M. Danev R. Nishiyama K. Sugawara K. & Nagayama K. In press. Zernike phase contrast electron microscopy of ice-embedded influenza A virus. J. Struct. Biol .
10.1016/S0031-8914(42)80035-X

Dates

Type	When
Created	17 years, 5 months ago (March 13, 2008, 8:08 p.m.)
Deposited	1 year, 6 months ago (Feb. 23, 2024, 2:25 p.m.)
Indexed	3 weeks, 5 days ago (Aug. 2, 2025, 12:08 a.m.)
Issued	17 years, 5 months ago (March 13, 2008)
Published	17 years, 5 months ago (March 13, 2008)
Published Online	17 years, 5 months ago (March 13, 2008)
Published Print	17 years, 2 months ago (June 27, 2008)

Funders 0

None

BibTeX

@article{Nagayama_2008, title={Phase contrast electron microscopy: development of thin-film phase plates and biological applications}, volume={363}, ISSN={1471-2970}, url={http://dx.doi.org/10.1098/rstb.2008.2268}, DOI={10.1098/rstb.2008.2268}, number={1500}, journal={Philosophical Transactions of the Royal Society B: Biological Sciences}, publisher={The Royal Society}, author={Nagayama, Kuniaki and Danev, Radostin}, year={2008}, month=mar, pages={2153–2162} }

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