The Problem of Demineralisation in Thin Sections of Fully Calcified Bone

Boothroyd, B.

doi:10.1083/jcb.20.1.165

Cited by 125 publications

(33 citation statements)

References 9 publications

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“…Washing of samples with dierent buers (Coetzee and Van-der-Merwe 1984), dehydration, and epoxy embedding (Mollenhauwer 1993) have also been reported as sources of artifacts during sample preparation for transmission electron microscopy. Likewise, sectioning (Boothroyd 1964;Harvey et al 1976;Morgan 1980) and staining with uranyl acetate (Agostini and Hasselbach 1971;Yarom et al 1974) can also lead to the loss of considerable amounts of some elements, mainly Na + and Ca 2+ . For that reason it seems that the appearance of the acidocalcisome in thin sections of epoxide-embedded cells (as an empty vacuole with only a thin layer of dense material closely apposed to the inner face of the membrane) probably represents an artifact produced after ®xation by the successive steps of washing, dehydration, and embedding during the sample preparation.…”

Section: Discussionmentioning

confidence: 98%

The fine structure of acidocalcisomes in Trypanosoma cruzi

Miranda¹,

Benchimol

Docampo

et al. 2000

Parasitology Research

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Trypanosoma cruzi survives in vertebrate and invertebrate hosts and has developed mechanisms that allow it to adapt to changes in the microenvironment such as temperature, pH, and ionic composition. Most of its calcium is concentrated in an organelle named the acidocalcisome, which is acidified by a (V-H+)-adenosine triphosphatase and has H+/Ca2+ counter-transportation for calcium uptake. In this work, acidocalcisomes were examined using different transmission electron microscopy techniques. In thin sections of different stages, acidocalcisomes presented a circular shape with an electron-dense inclusion containing P3-, Ca2+, Na+, Mg2+, K+, and Zn2+. They could be distinguished from gold-labeled albumin-containing reservosomes in whole epimastigotes, and a morphometric analysis showed higher amounts of these organelles in amastigotes as compared with epimastigotes and trypomastigotes. It is possible that this variation in the amount of acidocalcisomes in the different evolutive stages could reflect adaptation mechanisms used by the parasite to survive and multiply in different environmental conditions.

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Section: Discussionmentioning

confidence: 98%

The fine structure of acidocalcisomes in Trypanosoma cruzi

Miranda¹,

Benchimol

Docampo

et al. 2000

Parasitology Research

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“…The present study has been extended further by analyzing 35 OI bone specimens and 25 age-and sitematched normal controls. Furthermore, cryoprocessing avoids artifactual demineralization that may possibly occur during tissue processing [18][19][20][21] and ultramicrotomy [22]. This is the first report on Ca/P ratios determined on cryosections of OI and normal bone, and the first one to express them separately according to clinical types.…”

Section: Discussionmentioning

confidence: 99%

Abnormal Mineral Composition of Osteogenesis Imperfecta Bone as Determined by Electron Probe X-ray Microanalysis on Conventional and Cryosections

1999

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Osteogenesis imperfecta (OI) is a genetic disorder of the connective tissue characterized by frequent bone fractures. The cause of bone fragility is still unknown even though substantial work on collagen has been done. We measured the calcium to phosphorus ratio (Ca/P) of bone mineral from 35 OI bone samples and 25 age- and site-matched control specimens, using electron probe X-ray microanalysis in the transmission electron microscope. Ultra-thin cryosections and conventionally prepared resin sections were used. Cryo-ultramicrotomy avoids any possible artifactual demineralization that may occur in conventional aqueous media. The Ca/P ratio obtained by these two methods was compared and there was no statistical difference between them. The results were differentiated according to the clinical types of OI for the first time. The Ca/P ratio of OI bone mineral was lower than normal in both resin and cryosections, and mirrored the severity of the disease. OI type II had the lowest ratio (Ca/P = 1.49) compared with normal age- and site-matched controls (Ca/P = 1.69). This abnormal mineral composition in OI type II could be a contributory factor to bone fragility in OI bone.

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“…A soft X-ray photograph of 60 Sato/Ishikawa/Shimada/Ezure/Salo Development of Human Temporomandibular Joint sections of a specimen reveals the precise level of calcifica tion when the photograph is examined with an image ana lyzer (Shimada et al, 1992). The problem with using recon structions of serial sections is that crystals of calcium apa tite are leached out during preparation of tissue for ultrastructural studies [Boothroyd, 1964;Landis et ah, 19771. This problem is accumulated when one analysis sections of calcified hard tissue at early developmental stage by using sectioned specimens with low levels of calcification. There fore, all specimens used in this study were prepared by a nonsectioning method.…”

Section: Discussionmentioning

confidence: 99%

Morphology and Analysis of the Development of the Human Temporomandibular Joint and Masticatory Muscle

Sato¹,

Ishikawa

Shimada

et al. 1994

Cells Tissues Organs

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The calcification levels of the mandible and the temporal bone of human fetuses, which ranged from 12 to 32 weeks of gestation, were systematically investigated with a soft X-ray analyzer linked to an image analyzer. The profile of the condylar process (head) revealed high levels of calcification, in contrast to that in the mandibular fossa of the temporal bone. The basal portion of the condylar process and the mandibular notch exhibited moderate calcification from 12 weeks of gestation. The weight and the cross-sectional areas of the muscle and the muscle fibers in masticatory muscles (masseter, temporal, medial, and lateral pterygoid muscles) are all increased gradually during development from 12 to 32 weeks of gestation. These changes in calcification and in cross-sectional area of muscle suggest that muscle development may be related to bone calcification during formation of the mandible.

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The Problem of Demineralisation in Thin Sections of Fully Calcified Bone

Cited by 125 publications

References 9 publications

The fine structure of acidocalcisomes in Trypanosoma cruzi

The fine structure of acidocalcisomes in Trypanosoma cruzi

Abnormal Mineral Composition of Osteogenesis Imperfecta Bone as Determined by Electron Probe X-ray Microanalysis on Conventional and Cryosections

Morphology and Analysis of the Development of the Human Temporomandibular Joint and Masticatory Muscle

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