Ultrasound-accelerated tissue formalin fixation and paraffin embedding

描述（由申请人提供）：肿瘤治疗的第一步是确保及时准确的病理诊断。福尔马林固定石蜡包埋（FFPE）已成为标准的组织保存方法，用于90%以上的临床组织学诊断。虽然它提供了优越的形态学和易于长期储存的临床标本，FFPE是耗时的，并不能完全支持当前的分子分析。我们研究的长期目标是将现代技术应用于医疗实践和病理诊断，以有效地对抗癌症和其他疾病。我们计划进一步发展超声加速组织保存（UTP）技术，并研究超声（US）促进福尔马林固定的机制。我们的具体目标是：(1)开发一种多组织保存处理器，带有可选的实时数字系统来监测和标准化组织固定水平；(2)通过对组织病理学、大分子完整性及其长期稳定性进行更多的统计评估来验证UTP技术；(3)开展机制研究，阐明含US和不含US的福尔马林固定对交联形成、酶活性和蛋白质构象变化的影响。我们已经证明，与传统FFPE相比，UTP在组织形态上提供了相似的保存，具有相似的长期储存稳定性，改善了蛋白质结构、抗原特性和mRNA完整性的保存。UTP允许基于UTP固定组织提取物的简单一般分子分析和分析。UTP也为通过调节超声时间和强度来控制和监测固定水平提供了良好的机会。我们假设，us促进的福尔马林固定将大大加速甲醛诱导的组织中的大分子交联，并“冻结”大分子及其构象，因为加速了固定反应。由于组织保存仍然是组织学诊断前的标准和一般要求，因此这项创新将对经济和公共卫生产生重大影响。

DESCRIPTION (provided by applicant): The first crucial step in cancer management is to assure timely and accurate pathological diagnoses. Formalin fixation and paraffin embedding (FFPE) has been a standard tissue preservation method employed in over 90% cases for clinical histology diagnosis. Though it provides superior morphology and easy long- term storage of clinical specimens, FFPE is time-consuming and does not fully support current molecular analyses. The long-term goal of our research is to apply modern techniques to medical practice and pathological diagnosis to effectively fight cancers and other diseases. Our proposed project is to further develop the ultrasound-accelerated tissue preservation (UTP) technology for multiple tissues and to study the mechanism of ultrasound (US)-facilitated formalin fixation. Our specific aims are: (1) to develop a multi- tissue preservation processor with an optional real-time digital system to monitor and standardize tissue fixation level; (2)to validate the UTP techniques by performing more statistical assessments on histopathology, macromolecule integrity, and their long-term stability; and (3) to conduct mechanism studies to elucidate the effects of formalin fixation with and without US on the formation of cross-linking, enzymatic activity, and protein conformational changes. We have demonstrated that in comparison to conventional FFPE, UTP provides similar preservation in tissue morphology with similar long-term storage stability, improved preservation of protein structure, antigen properties, and mRNA integrity. UTP allows easy general molecular profiling and analyses based on extracts from UTP-fixed tissues. UTP also provides a good opportunity to control and monitor fixation level by adjusting the time and strength of the ultrasound. We hypothesize that US-facilitated formalin fixation will greatly accelerate formaldehyde-induced macromolecule cross-linking in tissues and "freeze" macromolecules and their conformation due to accelerated fixation reactions. Since tissue preservation is still a standard and general requirement before histology diagnosis, the innovation should have great impact in economy and public health.