The Experts below are selected from a list of 99 Experts worldwide ranked by ideXlab platform
Magdi M Salih - One of the best experts on this subject based on the ideXlab platform.
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comparison between conventional Decalcification and a microwave assisted method in Bone tissue affected with mycetoma
Biochemistry Research International, 2020Co-Authors: Magdi M SalihAbstract:Mycetoma is a lifelong granulomatous disease of subcutaneous tissues and Bones. Histopathology is a substantiated indicative method based on the assumption of a definitive diagnosis of mycetoma. It requires efficient processing of tissues including Bone Decalcification. The Decalcification process must ensure complete removal of calcium and also a proper preservation of tissue and microorganisms' staining ability. Objectives. To compare the conventional method used in Decalcification with the microwave method using different Decalcification solutions. Different characteristics were tested, including the speed of Decalcification and morphological and fungal preservation in Bone tissue affected with mycetoma. Materials and Methods. Three Decalcification solutions were employed to remove calcium from 50 Bone tissue samples affected with mycetoma, including 10% neutral buffered EDTA (pH 7.4), 5% nitric acid, and 5% hydrochloric acid. Conventional and microwave methods were used. Haematoxylin-eosin (HE) stain, Gridley's stain, and Grocott hexamine-silver stain were employed to evaluate the Bone and fungi morphologies. Results. The Decalcification time of the conventional method compared with the microwave method with 10% EDTA (pH 7.4) took 120 hours and 29 hours, while 5% hydrochloric acid and 5% nitric acid took 8 hours and 3 hours, separately. Also, 10% EDTA is the best decalcifying agent for HE staining and fungal stains. 5% hydrochloric acid and 5% nitric acid can be used for fungal staining. Conclusion. The current study investigated the effects of different decalcifying agents as well as two Decalcification procedures on the preservation of the Bone structure and fungal staining, which will help to develop suitable protocols for the analyses of the Bone tissue affected with mycetoma infection.
Kazuhiko Imaizumi - One of the best experts on this subject based on the ideXlab platform.
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application of microwave irradiation to Bone Decalcification and its effect on dna quality
2017Co-Authors: Kazuhiko ImaizumiAbstract:Microwave (MW) irradiation has been widely used in the field of histopathology to reduce the total time to prepare tissue sections for microscopy. The processes before tissue sectioning include fixation, dehydration, and embedding in paraffin, and MW irradiation accelerates these reactions. In histological studies of Bone, a process to soften compact Bone, “Decalcification”, is necessary. Many studies have applied MW irradiation to this time-consuming process, and positive results of MW irradiation for Bone Decalcification are accumulating. In recent years, genetic analysis has become popular in this field through the techniques of in situ hybridization and reverse transcription-polymerase chain reaction as well as genotyping of DNA extracted from decalcified specimens. The effect of MW irradiation on the quality of nucleic acids is now a major consideration. This chapter presents the MW effects on Bone Decalcification in terms of Bone histology, the progress of Decalcification techniques based on MW, and trends concerning the applicability of specimens decalcified with MWs for genetic analysis.
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an evaluation of the effect of microwave irradiation on Bone Decalcification aimed to dna extraction
Legal Medicine, 2013Co-Authors: Kazuhiko Imaizumi, Kei Taniguchi, Yoshinori OgawaAbstract:An effect of intermittent microwave irradiation on Decalcification of compact Bone followed by DNA extraction was verified. In order to perform quantitative analysis regarding the degree of Decalcification, Cubic Bone specimens were prepared from bovine metacarpal Bone and micro-focus X-ray CT imaging was applied to measure precise volume of decalcified area in the cubes. Microwave irradiation was performed under strict control of temperature using commercially available experimental device which is designed for advancing tissue fixation, Decalcification, and antigen-antibody reaction by intermittent microwave. The integrity of the DNA obtained from irradiated specimen was also examined by PCR analysis. The results of morphological analysis with CT imaging showed that microwave irradiation has a positive effect on Decalcification though that effect is not so drastic. The results obtained from PCR analysis showed that microwave irradiation decrease amplifiable DNA, suggesting that we should be careful to use microwave for the purpose of Bone DNA extraction.
Herman Van Dekken - One of the best experts on this subject based on the ideXlab platform.
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effect of Bone Decalcification procedures on dna in situ hybridization and comparative genomic hybridization edta is highly preferable to a routinely used acid decalcifier
Journal of Histochemistry and Cytochemistry, 1999Co-Authors: Janneke C Alers, Pieterjaap Krijtenburg, Kees J Vissers, Herman Van DekkenAbstract:Decalcification is routinely performed for histological studies of Bone-containing tissue. Although DNA in situ hybridization (ISH) and comparative genomic hybridization (CGH) have been successfully employed on archival material, little has been reported on the use of these techniques on archival decalcified bony material. In this study we compared the effects of two commonly used decalcifiers, i.e., one proprietary, acid-based agent (RDO) and one chelating agent (EDTA), in relation to subsequent DNA ISH and CGH to bony tissues (two normal vertebrae, six prostate tumor Bone metastases with one sample decalcified by both EDTA and RDO). We found that RDO-decalcified tissue was not suited for DNA ISH in tissue sections with centromere-specific probes, whereas we were able to adequately determine the chromosomal status of EDTA-decalcified material of both control and tumor material. Gel electrophoresis revealed that no DNA could be successfully retrieved from RDO-treated material. Moreover, in contrast to RDO...
Ccv Eloka - One of the best experts on this subject based on the ideXlab platform.
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a comparative study on the rate of Bone Decalcification at varying temperatures fluid type and fluid concentrations
International Journal of Basic Applied and Innovative Research, 2012Co-Authors: J K Bankole, Ccv Eloka, C J Okoro, R A Anyanwa, M Uwigbe, D OsiagwuAbstract:This study evaluates the rate of Bone Decalcification at varying temperatures, Decalcification fluid and fluid concentrations. A Decalcification methodology was adopted using modern household microwave oven to accelerate the Decalcification rate of rabbit compact Bone sample. Bone biopsy was obtained from a rabbit limb and fixed immediately for 24hrs in 10% formalin. Traditional Decalcification was carried out at room temperature (RT) with 5% Jenkins fluid (JK) and 5% Gooding and Stewart (GS) fluid as controls. Microwave oven Decalcification with 5% JK and 7% JK, as well as 5% GS and 10% GS served as the tests at 30 0 C, 40 0 C and 50 0 C in microwave oven. Results showed that at 30 0 C, 5%-GS and 10%-GS as well as 5%-JK and 7%-JK presented a decalcifying time of 11hrs:30mins and 10hrs:30mins as well as 15hrs and 13hrs respectively. At 40 0 C, a decalcifying time of 8hrs, 6hrs:30mins, 10hrs:30mins and 12hrs were recorded for the respective concentrations of decalcifying fluid. Also, at 50 0 C, the decalcifying times were 6hrs, 5hrs, and 9hrs:30mins and 8hrs respectively. The observed differences between RT and microwave oven Decalcification was significant while the rate between the two decalcifying fluid were not. Our findings further encourage the use of microwave oven for Bone Decalcification. Key Words: Bone, Decalcification rate, fluid concentration, Temperature
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the histological effects of microwave oven on Bone Decalcification using gooding and stewart fluid at varying temperatures and concentrations
International Journal of Basic Applied and Innovative Research, 2012Co-Authors: Ccv Eloka, C J Okoro, A C Okon, A P Eneasato, S N Ekoh, Gac EzeahAbstract:This study evaluates the histological preservation of Bone tissue architecture at varying temperatures and concentrations of Gooding and Stewart (GS) decalcifying fluid as the preservation of tissue architecture depends on the quality and velocity of the Decalcification processes. In the present study, a Decalcification methodology was adopted using household microwave oven to induce the Decalcification rate of rabbit compact Bone sample for microscopic analysis. Bone biopsy was obtained from rabbit limb and fixed immediately for 24hrs in 10% formalin. Traditional Decalcification was carried out at RT (Room Temperature) with 5%-GS as control while microwave oven Decalcification was conducted with 5%-GS and 10%-GS at 30 0 C, 40 0 C and 50 0 C respectively. The results showed that 5%-GS and 10%-GS gave excellent histological architecture at 30 0 C while at 40 0 C, the histological architectures were comparable to RT Decalcification. But at 50 0 C, Bone tissue architecture was completely destroyed being ghost-like in appearance. Of particular interest is the fact that the Bone tissue treated with 5%-GS irrespective of the temperature presented excellent tissue architecture than that of 10%-GS. Thus 5%-GS gave the best result in this study. Key Words: Microwave Oven, Decalcification, Decalcifying fluid, Tissue architecture.
Yoshinori Ogawa - One of the best experts on this subject based on the ideXlab platform.
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an evaluation of the effect of microwave irradiation on Bone Decalcification aimed to dna extraction
Legal Medicine, 2013Co-Authors: Kazuhiko Imaizumi, Kei Taniguchi, Yoshinori OgawaAbstract:An effect of intermittent microwave irradiation on Decalcification of compact Bone followed by DNA extraction was verified. In order to perform quantitative analysis regarding the degree of Decalcification, Cubic Bone specimens were prepared from bovine metacarpal Bone and micro-focus X-ray CT imaging was applied to measure precise volume of decalcified area in the cubes. Microwave irradiation was performed under strict control of temperature using commercially available experimental device which is designed for advancing tissue fixation, Decalcification, and antigen-antibody reaction by intermittent microwave. The integrity of the DNA obtained from irradiated specimen was also examined by PCR analysis. The results of morphological analysis with CT imaging showed that microwave irradiation has a positive effect on Decalcification though that effect is not so drastic. The results obtained from PCR analysis showed that microwave irradiation decrease amplifiable DNA, suggesting that we should be careful to use microwave for the purpose of Bone DNA extraction.