Lei Zhubing, Pang Xinpei, Li Li, Zhang Fan, Dong Wen-Fei, Mei Qian
School of Biomedical Engineering (Suzhou), Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, 230026, China.
CAS Key Laboratory of Biomedical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences (CAS), Suzhou, 215163, China.
Anal Bioanal Chem. 2022 Dec;414(29-30):8201-8213. doi: 10.1007/s00216-022-04360-5. Epub 2022 Oct 19.
Formalin-fixed paraffin-embedded (FFPE) tissue samples are routinely used in prospective and retrospective studies. It is crucial to obtain high-quality nucleic acid (NA) from FFPE samples for downstream molecular analysis, such as quantitative polymerase chain reaction (PCR), Sanger sequencing, next-generation sequencing, and microarray, in both clinical diagnosis and basic research. The current NA extraction methods from FFPE samples using chemical solvent are tedious, environmentally unfriendly, and unamenable to automation or field deployment. We present a tool for NA extraction from FFPE samples using a high-intensity focused ultrasound (HIFU) technology. A cartridge strip containing reagents for FFPE sample deparaffinization and NA extraction and purification is operated by an automation tool consisting of a HIFU module, a liquid handling robot unit, and accessories including a thermal block and magnets. The HIFU module is a single concaved piezoelectric ceramic plate driven by a current-mode class-D power amplifier. Based on the ultrasonic cavitation effects, the HIFU module provides highly concentrated energy introducing paraffin emulsification and disintegration. The high quantity and quality of NA extracted using the reported system are evaluated by PCR and compared with the quantity and quality of NA extracted using the current standard methods.
福尔马林固定石蜡包埋（FFPE）组织样本常用于前瞻性和回顾性研究。从FFPE样本中获取高质量核酸（NA）对于下游分子分析至关重要，这些分析包括临床诊断和基础研究中的定量聚合酶链反应（PCR）、桑格测序、新一代测序以及微阵列分析等。目前使用化学溶剂从FFPE样本中提取NA的方法繁琐、不环保，且不适用于自动化或现场部署。我们展示了一种使用高强度聚焦超声（HIFU）技术从FFPE样本中提取NA的工具。一个包含用于FFPE样本脱石蜡以及NA提取和纯化试剂的试剂盒条，由一个由HIFU模块、液体处理机器人单元以及包括热块和磁体在内的附件组成的自动化工具操作。HIFU模块是一个由电流模式D类功率放大器驱动的单凹压电陶瓷板。基于超声空化效应，HIFU模块提供高度集中的能量，使石蜡乳化和分解。使用报道的系统提取的NA的量和质量通过PCR进行评估，并与使用当前标准方法提取的NA的量和质量进行比较。
