Volume 1

Contemporary classics in the life sciences / edited by James T. Barrett.

Date:
©1986-
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Licence: Attribution-NonCommercial 4.0 International (CC BY-NC 4.0)

Credit: Contemporary classics in the life sciences / edited by James T. Barrett. Source: Wellcome Collection.

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    Number 47 Citation Classics November 20, 1978 Steere R.L. Electron microscopy of structural detail in frozen biological specimens. J. Biophys. Biochem. Cytol. 3:45-60, 1957. This paper describes a procedure for the preparation of pre-shadowed replicas which reveal fine ultrastructural details of frozen fractured, etched (by vacuum sublimation) biological specimens. Il lustrations reveal the ordered arrays of plant virus particles within crystals, in cluding tobacco mosaic virus crystals within infected cells. [The SCI® in dicates that this paper was cited 185 times in the period 1961-1977.] Russell L. Steere Plant Virology Laboratory Plant Protection Institute Beltsville Agricultural Research Center US Department of Agriculture Beltsville, MD 20705 January 13,1978 I believe my pleasure in finding this report in the 'Citation Classics' list results from its appearance as the first successful application of an alternative approach (freeze-etching) to the visualization of biological ultrastructure by electron microscopy. This approach came about as a logical consequence of learned techniques, equipment, and information available to me at the time, and complete freedom to ex plore the unknown mixed with unsatiated curiosity, eternal optimism, and Steere stub bornness! The work was done under the supervi sion of Robley C. Williams in the virus laboratory at the University of California, Berkeley, where we had just demonstrated that the beautiful crystals found in cells of tobacco mosaic virus (TMV>infected plants were composed V» of TMV particles and '/» of a volatile matrix (probably water). We knew that chemical fixation destroyed these extremely fragile crystals so that thin sec tion electron microscopy could not reveal their internal particle orientation. When I discussed the approach des cribed in this paper with Robley in 1952 as a means of visualizing the true crystal struc ture, he remarked that it would hardly be worth the 3 to 4 years it might take to develop such a procedure just to visualize the ultrastructure of TMV crystals but that, as an alternative procedure for studying cytological ultrastructure, it might be ex tremely worthwhile. He cautioned me that he couldn't guarantee a promotion and that I might spend 3 to 4 years on such a develop ment, then have to abandon it because of insurmountable obstacles. However, he did promise me that he would guarantee my job for at least 4 years if I wanted to tackle this important work. Now, after 20 years and numerous modifications to equipment and proce dures, freeze-etching and related techniques have finally come of age. This approach to the study of biological ultrastructure has not only revealed the finite structure of TMV crystals and confirmed many results obtained by thin sectioning and other techniques, but has provided much visual information on membrane and other ultrastructural details unobtainable as yet by other approaches. Recent improvements have pushed the resolution of freeze- etching to the range of 1 nm, and permit the comparison of complementary faces of frozen fractured specimens. While doing the research leading to the publication of this paper, I had the rare pleasure of working under exceptional research administrators, whose main con cerns were obtaining adequate funding for their programs, selecting qualified scien tists, giving them the freedom to explore the unknown, and providing them with ade quate protection from the 'publish or perish' philosophy, unnecessary 'busy work,' report writing, meetings, and other distractions.