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Results for “Decalcification Technique”

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At least 19 recordsLinked to original sources
PubMed1986

A technique for decalcification and demonstration of substance P-like immunoreactivity.

Substance P-like immunoreactivity (SPLI) was demonstrated in mouse spinal cord by an indirect immunofluorescence method after decalcification of the vertebra with a mixture of EDTA and Zamboni's fixative. SPLI was observed mainly in the gray matter of the spinal cord, especially the superficial layers of the dorsal horn; the distribution was the same as in the control spinal cord. No diffusion and depletion of SPLI were recognized after decalcification and no specific fluorescence was observed. The findings reported here indicate that decalcification with a mixture of EDTA and Zamboni's fixative is a useful method for examining SPLI in nervous tissue surrounded in situ by calcified tissues.

Animals↗
PubMed1991

Decalcification by perfusion. A new method for rapid softening of temporal bones.

We describe a new technique, decalcification by perfusion, for the softening of bony tissue. The blood circulatory system was perfused in 16 rats via a cannula through the left heart ventricle with a fixative followed by New Decalc (an acidic demineralizer) for 30-240 minutes. Perfusion decalcification for 120 minutes softened all heads and middle ear specimens could be easily sampled and prepared for studies by both light and electron microscope. For comparison, a conventional immersion technique required 72 hours of decalcification to accomplish softening. The perfusion technique considerably reduced the time needed to decalcify the tissue and preserved the morphology better than did the immersion procedure.

Animals↗
PubMedDate not supplied

[Focal demineralization of the cortical plate of the jaw and its experimental reversibility based on microphotometry data].

Focal demineralization of bone tissue with local dosed vacuum (exposure of up to 20 sec) and 2% lithium chloride electrophoresis (8 sessions daily, 10 min exposure) was tried in 40 white rats (21 reference ones and 19 experimental). The maximal reduction of mineral content was achieved on days 3-7 after the end of the course and made up 76.2 +/- 15.0% (p less than 0.01) and 80.6 +/- 12.5% (p less than 0.001), respectively, as against the reference animals. Mineral content at the site of demineralization gradually normalized by day 21. This transient reduction of mineral content of bone tissue may be used in orthodontic treatment of dentition abnormalities.

Alveolar Process↗
PubMed1991

Addition of cetyl pyridinium chloride to histological demineralizing agents to improve the demonstrability of gram-positive bacteria.

It has previously been shown experimentally using S. faecalis that the relative number of Gram-positive bacteria is reduced by demineralizing agents. To achieve better bacterial preservation after demineralization, cetyl pyridinium chloride (CPC) and neutral formaldehyde solution (NF) were added, separately and in combination, to formic acid and EDTA. The number of S. faecalis organisms in suspension was determined using a haemocytometer, and the percentage of Gram-positive bacteria was ascertained from smears. Both CPC and NF accomplished better bacterial preservation; when both substances were added together preservation was improved still further. The best results were achieved when CPC and NF were added to EDTA. If these findings could be extrapolated to tissue sections, and if it were intended to demonstrate Gram-positive bacteria, 10 per cent EDTA to which 0.5 per cent CPC and 4 per cent NF had been added would be recommended for demineralization.

Bacteriological Techniques↗
PubMed1978

Fluoride-induced mineralization within vacuoles in maturative ameloblasts of the rat.

Dense crystalline deposits appeared within vacuoles in rat maturative ameloblasts as a result of repeated injections of sodium fluoride. The crystals assumed varied arrangements but were always observed within intracellular vacuoles. The crystals resemble those of normal enamel and resist microincineration. They are readily dissolved by decalcification and leave behind an organic framework which matches the outline of the crystals. An experimental model is presented which may be useful in further studies of calcium transport, enamel matrix absorption or digestion of cellular debris.

Ameloblasts↗
PubMedDate not supplied

[Morphometric analysis of periosteocytic osteolysis: its application to the diagnosis of hyperparathyroidism].

The authors indicate the required methodologic conditions for the morphometric analysis of periosteocytic osteolysis, a reliable and specific feature of overactivity of the parathyroid gland. Indeed, significant periosteocytic enlargement has been found in 101 113 cases of primary hyperparathyroidism compared to 92 reference cases. The measurement of the size of the periosteocytic lacunae may be carried out either on hand of a micrometric eyepiece or of a picture analyser. It has to be carried out on decalcified bone sections of 5 microns thickness and it is necessary to measure at least 50 lacunae. The decalcification process unmasks an already partially decalcified crown of periosteocytic bone tissue the thickness of which is more important in the hyperparathyroidal bone than in normal bone. This zone has a peculiar collagen texture. The measurement of the peri-osteocytic enlargement is easy on stained sections of decalcified bone or on microradiographs of non-decalcified bone and, contrariwise, very dubious on stained sections of non-decalcified bone.

Automation↗
PubMed1991

The cumulative effect of disinfection, storage, histological fixation and demineralization on number and staining ability of gram-positive bacteria.

It has previously been shown experimentally using S. faecalis that the relative number of blue-staining Gram-positive bacteria was reduced by demineralizing agents as well as by disinfection (Wijnbergen & van Mullem 1987, van Mullem & Wijnbergen 1989). In this study the cumulative effect of disinfection, experimental period, fixation and demineralization was investigated. The use of additives to the fixative and the demineralizing agents to limit the loss of blue-staining bacteria was also studied. The numbers of bacteria were determined using a haemocytometer, and the percentage of blue-staining organisms were ascertained from smears. When compared with the start of the experiment, the sequence of disinfection with chlorhexidine, storage in PBS, fixation with neutral formaldehyde and demineralization with formic acid or EDTA appeared to reduce the relative numbers of Gram-positive staining bacteria. For storage periods of 0 and 4 days the reduction factors were 100 and 600, respectively, using formic acid. These factors were 50 and 95, respectively, using EDTA. Addition of cetyl pyridinium chloride to the fixative and the demineralizing agents, and addition of neutral formaldehyde to the demineralizing agents lowered these reduction factors to 80 and 200, respectively, for formic acid, and 40 and 85, respectively, for EDTA. If the results are extrapolated to animal experiments where disinfection, experimental period, fixation and demineralization form part of the experimental framework, even the lowest reduction in the number of blue-staining bacteria could lead to false interpretation of tissue sections.

Bacteriological Techniques↗
PubMed1975

Accelerated reattachment with cementogenesis to dentin, demineralized in situ. I. Optimum range.

This study confirms an original report describing accelerated reattachment with cementogenesis to root dentin, surgically exposed and demineralized in situ. It additionally describes results of 250 experiments on over 1000 teeth in mongrel dogs and cats designed to identify an optimum range of demineralization related to type of agent, pH, and time of application. This optimum range consistently induces flap reattachment with cementogenesis, while demineralization rates above and below this range enhance reattachment relative to undermineralized controls, but do not consistently induce new cementum. Although most controls demonstrated some reattachment with partial cementogenesis, none produced complete repair as did properly demineralized root surfaces and approximately one-third showed spithelial migration to the apical borders of the wound. No demineralized teeth demonstrated this control result. Small species differences in the response to root demineralization and the degree of hypermineralization of roots adjacent to chronic periodontal pockets, may make optimum ranges determined in this animal study slightly low for human pocket reaattachment techniques.

Animals↗
PubMed1978

Methods of preparation of fossil bone samples for light and transmission electron microscopy.

The methods described make possible the preparation of fossil samples for light nad transmission electron microscopy. To obtain good images with ground sections of bone, both the grinding process and classical bone staining techniques were modified, the latter by staining already made ground sections. Ground sections of suitable thickness were etched with a 4:1 mixture of 5% HNO3 and 2% OsO4 at a lowered temperature, thereby obtaining isolated osteocytes with a complete network of processes. Using the same method with thicker ground sections, the contents of vascular canals were isolated. The contents retained spatial relations permitting complete reconstruction of the vascular system of the bone in question. The isolated osteocytes, the vascular canal contents, and the isolated remnants of the intercellular organic part of this bone, subsequently embedded in Epon 812 and in methacrylate, made possible sections suitable for detailed analysis of the image of the surfaces of these elements. Polished surfaces of the fossil bone were replicated by the so-called clearing replicas technique adapted to fossil bone. The latter facilitated analysis of the ultrastructure of the surfaces of the vascular canals, isolated osteocytes and their processes, and analysis of the immediate proximity of these elements, since the technique preserves the mutual spatial relations intact.

Animals↗
PubMed1991

[Frame-mounted anastomosis of veins using a microsurgical technique and demineralized tubular allobone].

The experimental-morphological analysis covers the results of 62 operations for establishment of major vein anastomosis. In three series of experiments microsurgical suturing was used, while the control series employed the techniques of general surgery. In the microsurgery group, the best outcome (100 percent patency without anticoagulant administration) was achieved in cases when the venous anastomosis was protected by the demineralized tubular bone. Good outcomes were seen in cases with application of the original technique of mounting the venous anastomosis on rings of tubular bone. The advantages of microsurgical frame-mounted suturing of major veins over the general surgical techniques are demonstrated.

Anastomosis, Surgical↗
PubMed1988

Computer-assisted densitometric image analysis in periodontal radiography. A methodological study.

A videobased computer assisted densitometric image analysis (CADIA) system to quantify alveolar bone density changes on standardized dental radiographs was tested. An algorithm was used for grey level correction of a subsequent image to the baseline image. Quantitative information regarding positive and/or negative grey level changes were obtained automatically. Comparison of the ability of CADIA to detect surgically induced bone loss with interpretation of digital subtraction images and conventional radiographic interpretation revealed that CADIA was the most sensitive of the 3 methods, followed by interpretation of digital subtraction images which was considerably more sensitive than conventional radiographic interpretation. CADIA was capable of assessing differences in alveolar bone changes due to periodontal surgery between sites exposed to ostectomy/osteoplasty and control sites and sites exposed to periodontal surgery without ostectomy/osteoplasty. Finally, CADIA was capable of assessing differences in remodeling activity over 4-6 weeks after periodontal surgery between 45 surgical sites and 45 control sites. The system offers an objective method to quantitatively follow alveolar bone density changes over time and appears to be the most sensitive of previously described radiographic interpretation techniques.

Absorptiometry, Photon↗
PubMed1988

A new decalcifying technique for immunohistochemical studies of calcified tissue, especially applicable to cell surface marker demonstration.

We have developed a new decalcifying technique for use in light and electron microscopy studies utilizing immunohistochemical staining of calcified tissues. Specimens containing calcified tissues can be adequately decalcified at a pH of 7.1-7.4 and temperature of -5 degrees C, without freezing, by use of a solution containing EDTA, sodium hydroxide, and glycerol. In this study, Leu-2a, Leu-3a, Leu-4, Leu-7, Leu-12, Leu-14, Leu-M1, Leu-M2, Leu-M3, and HLA-DR-positive cells in destructive lesions of bone tissues from patients with rheumatoid arthritis and osteomyelitis were successfully detected immunohistochemically. Furthermore, the presence of HLA-DR antigen on the surface of the infiltrating cells in the same lesions could be demonstrated using the immunoelectron microscopic technique. The results reported here have not previously been obtainable using conventional decalcifying techniques.

Antibodies, Monoclonal↗
PubMed1992

The microwave oven in temporal bone research.

The microwave oven has recently been introduced in morphological research. It can be used for virtually all procedures in histological processing; fixation, dehydration, embedding, staining, immunotechniques, and the techniques used for electron microscopy. Here we report that the microwave oven histoprocedures also have several advantages in temporal bone research on animals. The microwave oven has been tested for (i) fixation, (ii) decalcification, (ii) staining of semithin plastic sections. The microwave oven procedures are time-saving and results in an excellent preservation of the tissues. The microwave histoprocessing should also be applicable to temporal bone research in human.

Animals↗
PubMed1990

Normal histology of the nasal cavity and application of special techniques.

There are three major epithelial types in the nasal mucosa, in addition to numerous accessory structures, some of which are species specific. Without careful and consistent processing of the nose tissue, histopathologic assessment of lesions in the nasal cavity may be compromised. While formalin fixation may be used for routine review of the nasal cavity, Bouin's fixation provides better histologic detail and fewer artifacts. Decalcification is not recommended for nasal tissues to be examined by transmission electron microscopy because of the detrimental effect of decalcifying solutions on sensory cells. Three levels of the nasal cavity may be used for routine histologic review of the nasal cavity, but four or five levels may be more appropriate for certain studies.

Acetates↗