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DOI 10.1007/s00216-003-1905-2
O R I G I N A L PA P E R
Received: 27 September 2002 / Revised: 3 March 2003 / Accepted: 11 March 2003 / Published online: 6 May 2003
Springer-Verlag 2003
Introduction
In nature, silica can be found in higher terrestrial plants
and a wide variety of organisms like diatoms, sponges and
molluscs [1, 2]. De Saussure (in Ref. [3]) first described
the silicification in plants. Silica distribution in various
grasses [4, 5, 6] has been widely studied and reported. The
shapes of phytoliths are characteristic of a given plant and
vary between different species [7, 8]. Usually silica is
found in parts of plants associated with cell wall in the
subterranean and aerial organs, those that infill the cell lumen and as extra cellular deposits. These silica deposits
can reproduce or mimic carbon-based cell structures in
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Fig. 1 (a) Cross section through the
stem of equisetum arvense; scale bar
200 m. (b) Silicon distribution in
Fig. 1a
Experimental
Materials and methods
Samples of shoots of Equisetum arvense were collected in autumn
and spring. Shoots were separated into stems and leaves. A part of
the sample was fixed with glutaraldehyde and subjected to critical
point drying so as to retain the cell structures without significant
deformation. A part of this fixed sample was further fixed with
OsO4 for contrasting between the organic part and silica and then
embedded in epoxy resin. Ultrathin sections of both longitudinal
and axial parts were made with a diamond knife on an ultramicrotome and placed on copper grids. These sections were viewed in a
LEO 912 TEM operated at 100 keV and electron diffraction was
made on the same instrument. TEM images and the diffraction pattern were recorded on image plates. Gold was used as the standard
for calibration. From the diffraction pattern of the samples and
gold standard, radial intensity scans were made. The scattering intensities were normalised and then Fourier transformed to difference atomic radial distribution functions [16, 17, 18]. Samples
were mounted on aluminium stubs and coated with gold for analysis by scanning electron microscopy. For elemental mapping, the
samples were taken as such without gold sputtering. In order to remove the organics for improved examination of the silica layer,
natural samples (without any fixation) were plasma ashed in an
oxygen atmosphere in a K1050 Plasma asher. The samples were
plasma ashed for 4.5 h with a power of 150 W. Plasma ashed and
natural samples were lightly ground with a pestle and mortar and
the powdered sample was subjected to SAXS studies. SAXS measurements were performed on a Kratky system using Ni-filtered
Cu K radiation (sealed tube, 2 kW X-ray generator); data were
collected and analysed by a previously described protocol [19].
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SAXS studies
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Fig. 11 EDX spectra of plasma ashed sample substantiating
the removal of organics
Conclusions
From the elemental mapping and EDX, we can conclude
that the silica is concentrated on the surface as a thin
layer. Ultra structure studies with TEM reveal the thickness of the surface layer to vary from a few nanometers to
about 7 m. The silica layer in the outer part was more
dense due to closer packed particles. The middle layer
also has a dense silica structure with some holes or chan-
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References
1. Kroger N, Deutzmann R, Sumper M (1999) Science 286:1129
1132
2. Kroger N, Deutzmann R, Sumper M (2001) Biol Chem 276:
2606626070
3. Jones LHP, Handreck KA (1967) Silica in soils plants and animals. Adv Agron 19:107149