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Secondary Ion Mass Spectrometry (SIMS) Microscopy as an Imaging Tool for Physiological Studies II. SIMS Microscopy of Plant Tissues

Published online by Cambridge University Press:  08 August 2003

Nicole Grignon
Affiliation:
Biochimie et Physiologic Végétales, INRA, Ecole Nationale Supérieure Agronomique, CNRS URA 573, Montpellier, France
Sylvain Halpern
Affiliation:
INSERM, Equipe de microscopie ionique, Institut Gustave-Roussy, Villejuif, France
Josette Jeusset
Affiliation:
INSERM, Equipe de microscopie ionique, Institut Gustave-Roussy, Villejuif, France
Philippe Fragu
Affiliation:
INSERM, Equipe de microscopie ionique, Institut Gustave-Roussy, Villejuif, France
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Abstract

Serious difficulties are encountered when SIMS analysis is applied to plant cells because of the cells' basic organization. In most plant cells, the cytoplasm is present as a thin layer that surrounds a large central vacuole, and is surrounded externally by a porous semi-rigid cell wall. Due to the high internal hydrostatic pressure typical of plant cells, large-scale solute redistribution may occur when tissues are excised. Relatively small solute decompartmentation is sufficient to collapse the native solute gradients between the cytoplasm and the adjacent compartments, due to the small volume of the former. For these reasons, most of the SIMS analyses in plant cells have been performed on elements bound to non-diffusible structures such as proteins, cell wall polymers, or in dry seeds. Sample preparation remains a limiting factor when imaging the distribution of soluble compounds. Cryotechniques have generated considerable interest to circumvent these problems. Cryofixation followed by cryosectioning would a priori be the best procedure, but encouraging results indicate that cryofixation followed by cryosubstitution is an interesting alternative.

Type
Research Article
Copyright
© 1996 Microscopy Society of America

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