Ultrasound-accelerated tissue formalin fixation and paraffin embedding

描述(由申请人提供)：癌症管理的第一个关键步骤是确保及时和准确的病理诊断。福尔马林固定石蜡包埋(FFPE)是一种标准的组织保存方法，90%以上的病例用于临床组织学诊断。尽管FFPE提供了优越的形态和易于长期保存的临床标本，但FFPE非常耗时，并不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。我们的计划是进一步发展超声加速多组织保存(UTP)技术，并研究超声促进福尔马林固定的机制。我们的具体目标是：(1)开发一种带有可选实时数字系统的多组织保存处理器，以监测和标准化组织固定水平；(2)通过对组织病理学、高分子完整性及其长期稳定性进行更多统计评估，验证UTP技术；以及(3)进行机制研究，以阐明使用和不使用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，与传统的FFPE相比，UTP在组织形态上提供了类似的保存，具有类似的长期保存稳定性，改善了蛋白质结构、抗原属性和mRNA完整性的保存。UTP允许基于UTP固定组织的提取物进行简单的一般分子分析和分析。UTP还提供了一个很好的机会，通过调整超声的时间和强度来控制和监测固定水平。我们假设，美国促进的福尔马林固定将极大地加速甲醛诱导的大分子在组织中的交联，并由于加速固定反应而“冻结”大分子及其构象。由于组织保存仍然是组织学诊断前的标准和普遍要求，这一创新应该会对经济和公众健康产生重大影响。

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.