Laboratory of Plastination and Anatomical Techniques, Centre for Research in Dental Sciences (CICO), Dental School, Universidad de La Frontera, Temuco, Chile; Center of Excellence in Morphological and Surgical Studies (CEMyQ), Medicine School, Universidad de La Frontera, Temuco, Chile; Doctoral Program in Morphological Sciences, Medicine School, Universidad de La Frontera, Temuco, Chile.
Laboratory of Plastination, Department of Medical Education, College of Medicine, University of Toledo, Ohio, USA.
Forensic Sci Int. 2020 Apr;309:110199. doi: 10.1016/j.forsciint.2020.110199. Epub 2020 Feb 15.
Plastination allows anatomical samples to be preserved in excellent condition for an indefinite period, free of formalin, and in a format that allows biosafe manipulation by students, academics, and researchers. As with other tissue preservation techniques, it is important to establish the level of conservation of deoxyribonucleic acid (DNA) for use in future applications. The object of the present work was to extract and evaluate DNA from plastinated tissues.
We used samples of liver from Canis lupus familiaris and skeletal muscle from Rattus norvegicus, Sprague-Dawley strain, extracted from specimens plastinated with silicone at room temperature. The tissue samples were deplastinated by incubation in 5% sodium methoxide dissolved in methanol for 24 or 48 h. The samples were divided into two equal parts and DNA was extracted using two different protocols. After extraction, the DNA was quantified by fluorometry and its integrity was assessed by electrophoresis in a 1% agarose gel.
A high yield of DNA was obtained from the deplastinated samples and the DNA was intact. Plastinated tissues have proven to be stable and easily managed. They can also be used for examination under light and electron microscopes. The DNA extraction technique used here allowed us to obtain intact DNA from samples plastinated with silicone at room temperature, without previous fixing. This technique may allow tissue specimens to be preserved for retrospective studies of archived samples of normal and pathological anatomy in the fields of basic, clinical, forensic, and epidemiological sciences.
The extracted DNA was intact and suitable for use in subsequent applications. Obtaining whole DNA from plastinated samples using tissue preservation protocols that preserve the tissue for use in subsequent applications, like real-time PCR, opens up many possibilities, with applications in the basic and clinical sciences, epidemiology, and forensic science.
塑化技术可使解剖标本在无福尔马林的情况下,以一种安全的方式被保存,并保持良好状态,保存时间也可无限延长。这种技术类似于其他组织保存技术,在用于未来应用之前,确定脱氧核糖核酸(DNA)的保存水平非常重要。本研究旨在从塑化组织中提取和评估 DNA。
我们使用犬科狼属的肝脏样本和 Sprague-Dawley 品系的大鼠骨骼肌样本,这些样本是从室温下用硅树脂塑化的标本中提取的。将组织样本在 5%甲醇钠的甲醇溶液中孵育 24 或 48 小时,使样本脱塑化。将样本分为两等份,并使用两种不同的方案提取 DNA。提取后,通过荧光光度法定量 DNA,通过 1%琼脂糖凝胶电泳评估其完整性。
从脱塑化的样本中获得了高产量的 DNA,并且 DNA 是完整的。事实证明,塑化组织稳定且易于处理。它们也可用于在光镜和电子显微镜下检查。这里使用的 DNA 提取技术使我们能够从室温下用硅树脂塑化的样本中获得完整的 DNA,而无需预先固定。这种技术可能允许保存组织标本,以便对基础、临床、法医和流行病学科学领域的正常和病理解剖存档样本进行回顾性研究。
提取的 DNA 是完整的,适用于后续应用。使用可保存组织以备后续应用(如实时 PCR)的组织保存方案从塑化样本中获得完整的 DNA,为基础和临床科学、流行病学和法医学开辟了许多可能性。