DOI: 10.1111/j.1365-2818.2006.01567.x. Focused ion beam milling of vitreous water: prospects for an alternative to cryo‐ultramicrotomy of frozen‐hydrated biological samples

Focused ion beam milling of vitreous water: prospects for an alternative to cryo‐ultramicrotomy of frozen‐hydrated biological samples

10.1111/j.1365-2818.2006.01567.x

Crossref journal-article

Wiley

Journal of Microscopy (311)

Abstract

SummaryThe feasibility of using a focused ion beam (FIB) for the purpose of thinning vitreously frozen biological specimens for transmission electron microscopy (TEM) was explored. A concern was whether heat transfer beyond the direct ion interaction layer might devitrify the ice. To test this possibility, we milled vitreously frozen water on a standard TEM grid with a 30‐keV Ga+ beam, and cryo‐transferred the grid to a TEM for examination. Following FIB milling of the vitreous ice from a thickness of approximately 1200 nm to 200–150 nm, changes characteristic of heat‐induced devitrification were not observed by TEM, in either images or diffraction patterns. Although numerous technical challenges remain, it is anticipated that ‘cryo‐FIB thinning’ of bulk frozen‐hydratred material will be capable of producing specimens for TEM cryo‐tomography with much greater efficiency than cryo‐ultramicrotomy, and without the specimen distortions and handling difficulties of the latter.

Bibliography

MARKO, M., HSIEH, C., MOBERLYCHAN, W., MANNELLA, C. A., & FRANK, J. (2006). Focused ion beam milling of vitreous water: prospects for an alternative to cryoâultramicrotomy of frozenâhydrated biological samples. Journal of Microscopy, 222(1), 42â47. Portico.

Authors 5

M. MARKO (first)
C. HSIEH (additional)
W. MOBERLYCHAN (additional)
C. A. MANNELLA (additional)
J. FRANK (additional)

References 22 Referenced 172

10.1046/j.1365-2818.2003.01244.x
10.1016/j.jsb.2005.01.003
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10.1007/978-1-4899-2302-8 / Low‐Temperature Microscopy and Microanalysis by Echlin P. (1992)
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{'issue': '2', 'key': 'e_1_2_6_12_1', 'first-page': '330CD', 'article-title': 'Cryo‐electron tomography and fluorescence microscopy of unicellular algae in vitreous cryosections', 'volume': '11', 'author': 'Leis A.', 'year': '2005', 'journal-title': 'Microsc. Microanal'} / Microsc. Microanal / Cryo‐electron tomography and fluorescence microscopy of unicellular algae in vitreous cryosections by Leis A. (2005)
10.1007/s11664-002-0169-5
10.1111/j.1365-2818.1991.tb03155.x
{'key': 'e_1_2_6_15_1', 'first-page': '8', 'article-title': 'The use of a SEM/FIB dual beam applied to biological samples', 'volume': '2', 'author': 'Mulders H.', 'year': '2003', 'journal-title': 'G.I.T. Imaging Microsc'} / G.I.T. Imaging Microsc / The use of a SEM/FIB dual beam applied to biological samples by Mulders H. (2003)
{'issue': '2', 'key': 'e_1_2_6_16_1', 'first-page': '150', 'article-title': 'Visualizing the macromolecular organization of chloroplast membranes using cryo‐electron tomography', 'volume': '11', 'author': 'Nicastro D.', 'year': '2005', 'journal-title': 'Microsc. Microanal'} / Microsc. Microanal / Visualizing the macromolecular organization of chloroplast membranes using cryo‐electron tomography by Nicastro D. (2005)
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10.1017/S1431927603445832 / Microsc. Microanal / Cryo‐electron tomography of frozen‐hydrated sections of eukaryotic cells by Schwartz C. (2003)
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10.1111/j.0022-2720.2004.01395.x
{'key': 'e_1_2_6_23_1', 'volume-title': 'The Stopping and Range of Ions in Solids', 'author': 'Ziegler J.F.', 'year': '1985'} / The Stopping and Range of Ions in Solids by Ziegler J.F. (1985)

Dates

Type	When
Created	19 years, 2 months ago (May 22, 2006, 11:47 a.m.)
Deposited	1 year, 10 months ago (Oct. 14, 2023, 10:25 p.m.)
Indexed	12 hours, 9 minutes ago (Aug. 20, 2025, 9:17 a.m.)
Issued	19 years, 4 months ago (April 1, 2006)
Published	19 years, 4 months ago (April 1, 2006)
Published Online	19 years, 2 months ago (May 22, 2006)
Published Print	19 years, 4 months ago (April 1, 2006)

Funders 0

None

BibTeX

@article{MARKO_2006, title={Focused ion beam milling of vitreous water: prospects for an alternative to cryo‐ultramicrotomy of frozen‐hydrated biological samples}, volume={222}, ISSN={1365-2818}, url={http://dx.doi.org/10.1111/j.1365-2818.2006.01567.x}, DOI={10.1111/j.1365-2818.2006.01567.x}, number={1}, journal={Journal of Microscopy}, publisher={Wiley}, author={MARKO, M. and HSIEH, C. and MOBERLYCHAN, W. and MANNELLA, C. A. and FRANK, J.}, year={2006}, month=apr, pages={42–47} }

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