Freeze substitution in 3 hours or less

McDonald, K. L. and Webb, R. I. (2011) Freeze substitution in 3 hours or less. Journal of Microscopy, 243 3: 227-233. doi:10.1111/j.1365-2818.2011.03526.x


Author McDonald, K. L.
Webb, R. I.
Title Freeze substitution in 3 hours or less
Journal name Journal of Microscopy   Check publisher's open access policy
ISSN 0022-2720
1365-2818
Publication date 2011-09
Sub-type Critical review of research, literature review, critical commentary
DOI 10.1111/j.1365-2818.2011.03526.x
Volume 243
Issue 3
Start page 227
End page 233
Total pages 7
Place of publication Oxford, United Kingdom
Publisher Wiley-Blackwell Publishing
Collection year 2012
Language eng
Formatted abstract
Summary
Freeze substitution is a process for low temperature dehydration and fixation of rapidly frozen cells that usually takes days to complete. New methods for freeze substitution have been developed that require only basic laboratory tools: a platform shaker, liquid nitrogen, a metal block with holes for cryotubes and an insulated container such as an ice bucket. With this equipment, excellent freeze substitution results can be obtained in as little as 90 min for cells of small volume such as bacteria and tissue culture cells. For cells of greater volume or that have significant diffusion barriers such as cuticles or thick cell walls, one can extend the time to 3 h or more with dry ice. The 3-h method works well for all manner of specimens, including plants and Caenorhabditis elegans as well as smaller samples. Here, we present the basics of the techniques and some results from Nicotiana leaves, C. elegans adult worms, Escherichia coli and baby hamster kidney tissue culture cells.
Keyword C. elegans
E. coli
Electron microscopy
High-pressure freezing
Freeze substitution
Tissue culture cells
Tobacco leaves
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Published under issue heading: "Hot Topic – fast-tracked short communication"

Document type: Journal Article
Sub-type: Critical review of research, literature review, critical commentary
Collections: Official 2012 Collection
Centre for Microscopy and Microanalysis Publications
 
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