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Imaging in head and neck surgery
Published in Neeraj Sethi, R. James A. England, Neil de Zoysa, Head, Neck and Thyroid Surgery, 2020
Both MR and CT can assess expansion into the larynx and cartilage destruction as well as parapharyngeal or paraglottic fat involvement. However, decalcification associated with increasing age must be taken into account when considering cartilage destruction.
Histological Study in Orthopaedic Animal Research
Published in Yuehuei H. An, Richard J. Friedman, Animal Models in Orthopaedic Research, 2020
Helen E. Gruber, Audrey A. Stasky
Decalcification procedures in general use either acids which react with the calcium in bone to form soluble calcium salts, or chelating agents which complex the calcium ions. Acid decalcifying agents are known to have the drawback of altering the staining properties of the embedded bone.14 Some decalcifying solutions also contain formalin, and thus increase the possibility of aldehyde groups increasing in the tissue and blocking reactions.
Head and Neck Pathology
Published in John C Watkinson, Raymond W Clarke, Terry M Jones, Vinidh Paleri, Nicholas White, Tim Woolford, Head & Neck Surgery Plastic Surgery, 2018
Ram Moorthy, Adrian T. Warfield, Max Robinson
After a further period of fixation, the tissue slices undergo cycles of dehydration, clearing, infiltration and embedding in preparation for microtomy (section cutting). During dehydration, alcohol replaces the aqueous fixative within the tissue. Clearing replaces the alcohol with an antemedium, such as xylene. Molten paraffin wax then replaces the clearing agent and infiltrates the tissue. The tissue is subsequently embedded by encapsulation in paraffin wax in a mould to provide a rigid support for microtomy. The additional step of decalcification may be instituted in mineralized tissue, which might otherwise hinder sectioning. Automatic tissue processors enhanced by pressure, vacuum, heat and microwave facilities in a self-contained, microprocessor-controlled, programmable unit are used in many modern laboratories, tailored to local conditions.
The effect of shock waves on mineralization and regeneration of distraction zone in osteoporotic rabbits
Published in Annals of Medicine, 2023
Enes Özkan, Erman Şenel, Mehmet Cihan Bereket, Mehmet Emin Önger
A blinded histologist conducted the preparation and stereological examination of tissue samples. Soft tissues of the jaws (skin, muscle, fascia, mucosa and periostium) were removed and were fixed in formaldehyde (10%) for one week. Following fixation, samples were exposed to decalcification in formic acid (5%) for 21 d. After decalcification, the tissues and gradually dehydrated with alcohol and were embedded in fresh paraffin. Serial sections of 7-μm thickness were taken from each paraffin block. Paraffin blocks were sampled at a ratio of 1/10 according to the volumetric estimation procedure, and the first section was randomly selected. Sections were stained with hematoxylin-eosin (HE) and photographed with a color digital camera using a light microscope in a stereology analysis system.
Diversity in matrilineages among the Jomon individuals of Japan
Published in Annals of Human Biology, 2023
Fuzuki Mizuno, Yasuhiro Taniguchi, Osamu Kondo, Michiko Hayashi, Kunihiko Kurosaki, Shintaroh Ueda
The powdered samples (100–500 mg) were decalcified in 0.5 M EDTA (pH 8.0) for 2 h at 56 °C in a rotating hybridisation oven, and the supernatant was removed by centrifugation. The decalcification step was performed thrice. DNA was extracted using phenol: chloroform: isoamyl alcohol (25:24:1) followed by extraction with an equal volume of chloroform. After centrifugation, the aqueous solution was removed and concentrated by centrifugation dialysis using the Amicon Ultra-15-30 kDa centrifugal filter (Merck Millipore) to a final volume of 200 μL. The DNA solution was purified using silica-based MiniElute spin columns (Qiagen), according to the manufacturer’s protocol. The obtained DNA was quantified using the Quant-iT dsDNA HS assay kit (Invitrogen), according to the manufacturer’s protocol. Using the DNA, we prepared single- and double-stranded NGS libraries. For the double-stranded libraries, we treated the DNA with PreCR Repair Mix (New England BioLabs). We carried out in-solution target enrichment using the SureSelect kit (Agilent Technologies), as described in a previous study (Kihana et al. 2013, Mizuno et al. 2020, 2021). The library was sequenced on the MiSeq platform (Illumina, USA) using the MiSeq Reagent Kit v3 150 cycles.
Isoorientin ameliorates osteoporosis and oxidative stress in postmenopausal rats
Published in Pharmaceutical Biology, 2022
Zhilin Cao, Wei Liu, Benjun Bi, Hao Wu, Gong Cheng, Zhongyuan Zhao
After fixing the femur for 24 h, decalcification was performed using 10% ethylenediaminetetraacetic acid (EDTA) for 30 days. The decalcification solution was replaced every 3 days, and decalcification of the bone tissue was checked during fluid exchange. The femoral tissue was placed in 95% ethanol overnight, gradient dehydrated to 100% ethanol for 1 h per gradient, xylene treated until transparent, embedded in paraffin, and sectioned into 6 μm. Sectional dewaxing with xylene was performed followed by gradient alcohol rehydration, distilled water soaking, haematoxylin (Solarbio, Beijing, China) staining for 20 min, washing in running water back to blue for 10 min, eosin (Solarbio, Beijing, China) staining for 10 min, gradient alcohol dehydration, xylene treatment until transparency, and neutral gum treatment for sealing. Pathological changes in the femoral tissue were observed under an optical microscope (magnification, ×100; Olympus, Japan). Image Pro Plus software (Media Cybernetics, USA) was used to calculate the percentage of the trabecular area (% Tb.Ar = trabecular area/total bone tissue area × 100).