Methods and Instrumentation for Hydrostatic Pressure-Enhanced Tissue Fixation

描述(申请人提供)：用福尔马林和石蜡固定组织样本是一种长期保存组织以供以后组织分析的既定方法。这些福尔马林固定的石蜡包埋组织也可以用于DNA、RNA或蛋白质的提取，用于追溯分子研究。不幸的是，传统的保存方法不能保证均匀的固定，经常导致标本外围分子的过度交联和深度的缓慢降解。因此，抗原回收、基因组或蛋白质组生物标志物鉴定研究可能会产生不准确或误导的结果。对流加热或微波辐射处理等替代固定方法的尝试也被证明是不够的，因为它们无法产生统一的产品，或导致生物分子轮廓的扰动。我们建议开发一种自动化的组织保存系统，该系统使用高静水压力来通过精确的热力学控制来标准化组织固定过程。高压已被证明可以增强甲醛快速和均匀地渗透到组织块中。拟议的组织保存系统将通过增强固定剂的渗透性来更好地保存组织，同时保存组织结构。这样的改进应该会导致从保存的样本中产生更好的数据，这可能会导致更快的疾病诊断和治疗方法的成功开发。1 与公众健康相关：拟议的研究旨在开发一种用于自动保存临床组织样本的高压系统，以促进和标准化组织固定过程，而不受样本大小和组织类型的影响。组织结构的更好、更一致和更均匀的保存，以及潜在生物标记物的化学完整性的改善，有望为使用固定样本库的未来组织病理学和回顾性分子诊断学研究带来可观的好处。

DESCRIPTION (provided by applicant): Fixation of tissue samples with formalin and paraffin is an established approach for long-term storage of tissues for later histological analyses. These formalin-fixed paraffin-embedded tissues can also be used for DNA, RNA or protein extraction for retrospective molecular studies. Unfortunately, traditional preservation methods do not guarantee uniform fixation, frequently resulting in excessive cross-linking of molecules on the periphery of the specimen and slow degradation in its depth. Consequently, antigen retrieval, genomic or proteomic biomarker identification studies may generate inaccurate or misleading results. Attempts at alternative fixation methods, such as convectional heat or microwave radiation treatment, have also proven inadequate, since they fail to produce a uniform product, or result in perturbations to the biomolecular profiles. We propose to develop an automated tissue preservation system that uses high hydrostatic pressure to standardize the process of tissue fixation with precise thermodynamic control. High pressure has been shown to enhance rapid and uniform formaldehyde penetration into tissue blocks. The proposed Tissue Preservation System will enable better preservation of tissues through enhanced penetration of fixative while preserving tissue architecture. Such improvements should lead to the generation of better data from preserved samples, which may lead to faster disease diagnosis and successful development of therapeutics. 1 PUBLIC HEALTH RELEVANCE: The proposed study aims to develop a high pressure system for automated preservation of clinical tissue specimens to facilitate and standardize the tissue fixation process independent of specimen size and tissue type. Better, more consistent, and more uniform preservation of tissue architecture, and improved chemical integrity of potential biomarkers, are expected to lead to considerable benefits for future histopathology and retrospective molecular diagnostics studies using fixed sample repositories.