An improved procedure for low-temperature embedding of high-pressure frozen and freeze-substituted plant tissues resulting in excellent structural preservation and contrast

Summary Here we describe refinements in the processing of high‐pressure frozen samples of delicate plant tissues for immuno‐electron microscopy. These involve: shortened freeze‐substitution schedules, lower temperatures during processing and polymerisation, the avoidance of temperature fluctuations...

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Bibliographic Details
Published in:Journal of microscopy (Oxford) 2012-07, Vol.247 (1), p.43-47
Main Authors: HILLMER, S., VIOTTI, C., ROBINSON, D.G.
Format: Article
Language:English
Subjects:
Aluminum
Botany - methods
Containers
Electron microscopy
Embedding
Freeze substitution
Freezing
Heat transfer
high-pressure freezing
Low temperature
low-temperature embedding
Microscopy - methods
Microscopy, Electron - methods
Plants - ultrastructure
Polymerization
Preservation
Pressure
Schedules
Specimen Handling - methods
Variations
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Summary:Summary Here we describe refinements in the processing of high‐pressure frozen samples of delicate plant tissues for immuno‐electron microscopy. These involve: shortened freeze‐substitution schedules, lower temperatures during processing and polymerisation, the avoidance of temperature fluctuations and the optimisation of heat transfer from the specimens using small disposable aluminium containers. The application of these modifications leads to very good structural preservation and selective membrane contrast. As a result, the versatility of the method is increased since not only immuno‐electron microscopical studies can be performed but often the quality is also quite suitable for structural investigations.
ISSN:0022-2720
1365-2818
DOI:10.1111/j.1365-2818.2011.03595.x