Comparison of the ultrastructure of conventionally fixed and high pressure frozen/freeze substituted root tips ofNicotiana andArabidopsis |
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Authors: | J Z Kiss Th H Giddings Jr L A Staehelin F D Sack |
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Institution: | (1) Department of Plant Biology, Ohio State University, Columbus, Ohio;(2) Department of Molecular, Cellular, and Developmental Biology, University of Colorado, 80309-0347 Boulder, CO, USA |
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Abstract: | Summary To circumvent the limitations of chemical fixation (CF) and to gain more reliable structural information about higher plant tissues, we have cryofixed root tips ofNicotiana andArabidopsis by high pressure freezing (HPF). Whereas other freezing techniques preserve tissue to a relatively shallow depth, HPF in conjunction with freeze substitution (FS) resulted in excellent preservation of entire root tips. Compared to CF, in tissue prepared by HPF/FS: (1) the plasmalemma and all internal membranes were much smoother and often coated on the cytoplasmic side by a thin layer of stained material, (2) the plasmalemma was appressed to the cell wall, (3) organelle profiles were rounder, (4) the cytoplasmic, mitochondrial, and amyloplast matrices were denser, (5) vacuoles contained electron dense material, (6) microtubules appeared to be more numerous and straighter, with crossbridges observed between them, (7) cisternae of endoplasmic reticulum (ER) were wider and filled with material, (8) Golgi intercisternal elements were more clearly resolved and were observed between both Golgi vesicles and cisternae, and (9) larger vesicles were associated with Golgi stacks. This study demonstrates that HPF/FS can be used to successfully preserve the ultrastructure of relatively large plant tissues without the use of intracellular cryoprotectants.Abbreviations CF
chemical fixation
- ER
endoplasmic reticulum
- FF
freeze fracture
- FS
freeze substitution
- HPF
high pressure freezing
Dedicated to the memory of Professor Oswald Kiermayer |
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Keywords: | Cryofixation Tobacco Ultrarapid freezing |
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