ISSN:
1615-6102
Keywords:
Actin
;
Field emission scanning electron microscope
;
High-resolution scanning electron microscopy
;
Immunolabelling
;
Microtubules
;
Plant cells
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Summary Two basic specimen preparation protocols that allow field emission scanning electron microscope imaging of intracellular structures in a wide range of plants are described. Both protocols depend on freeze fracturing to reveal areas of interest and selective removal of cytosol. Removal of cytosol was achieved either by macerating fixed tissues in a dilute solution of osmium tetroxide after freeze fracturing or by permeabilizing the membranes in saponin before fixation and subsequent freeze fracturing. Images of a variety of intracellular structures including all the main organelles as well as cytoskeletal components are presented. The permeabilization protocol can be combined with immunogold labelling to identify specific components such as microtubules. High-resolution three-dimensional imaging was combined with immunogold labelling of microtubules and actin cables in cell-free systems. This approach should be especially valuable for the study of dynamic cellular processes (such as cytoplasmic streaming) in live cells when used in conjunction with modern fluorescence microscopical techniques.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF01276854
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