Ultrasound-accelerated tissue formalin fixation and paraffin embedding

超声加速组织福尔马林固定和石蜡包埋

• 批准号: 7089318
• 负责人: Wei-Sing Chu
• 金额: $ 20.41万
• 依托单位: AMERICAN REGISTRY OF PATHOLOGY, INC.
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7089318
• 关键词: DNA; RNA; animal tissue; bioengineering /biomedical engineering; biomedical equipment development; buffers; cell morphology; clinical research; enzyme activity; free radicals; histochemistry /cytochemistry; human tissue; immunocytochemistry; in situ hybridization; polymerase chain reaction; proteins; stainings; tissue /cell preparation; tissue /organ preservation; ultrasound

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.

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准确的病理诊断。福尔马林固定石蜡包埋（FFPE）已成为90%以上病例的标准组织保存方法。尽管FFPE提供了上级的形态学和易于长期储存的临床标本，但它耗时并且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，以有效对抗癌症和其他疾病。本研究拟进一步发展超声加速组织保存技术，并探讨超声促进福尔马林固定的机制。我们的具体目标是：（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;以及（3）进行机理研究以阐明有和没有US的福尔马林固定对交联形成、酶活性和蛋白质构象变化。我们已经证明，与常规FFPE相比，UTP提供了类似的组织形态保存，具有类似的长期储存稳定性，改善了蛋白质结构、抗原性质和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谱分析和分析。UTP还提供了通过调整超声的时间和强度来控制和监测固视水平的良好机会。我们假设，美国促进福尔马林固定将大大加速甲醛诱导的大分子交联的组织和“冻结”大分子及其构象，由于加速固定反应。由于组织保存仍然是组织学诊断前的标准和一般要求，因此这一创新将在经济和公共卫生方面产生重大影响。