Methods and Instrumentation for Hydrostatic Pressure-Enhanced Tissue Fixation

静水压力增强组织固定的方法和仪器

• 批准号: 8201247
• 负责人: Alexander V Lazarev
• 金额: $ 16.1万
• 依托单位: PRESSURE BIOSCIENCE, INC.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2012-08-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8201247
• 关键词: Address; Affect; Antigens; Architecture; Bathing; Biological Markers; Biological Preservation; Cellular Morphology; Chemicals; Clinical; Cytolysis; DNA; Data; Dehydration; Diagnostic; Dimensions; Electron Microscopy; Ensure; Ethanol; Fixatives; Formaldehyde; Formalin; Frequencies; Future; Generations; Genomics; Glutaral; Heating; High temperature of physical object; Histocompatibility Testing; Histopathology; Hydration status; Hydrostatic Pressure; Imagery; Immunohistochemistry; Lead; Liquid substance; Magnetic Resonance Imaging; Methods; Microscopy; Modeling; Modification; Molecular; Monitor; Paraffin; Paraffin Embedding; Penetration; Performance; Perfusion; Phase; Preparation; Process; Proteins; Proteomics; Protocols documentation; Pulse Pressure; RNA; Reaction; Reagent; Reproducibility; Retrieval; Sampling; Series; Specimen; Speed; Staging; System; Technology; Temperature; Testing; Thermodynamics; Time; Tissue Fixation; Tissue Preservation; Tissue Sample; Tissues; Water; Xylene; aqueous; base; carbene; crosslink; design; disease diagnosis; flexibility; improved; instrument; instrumentation; meetings; microwave electromagnetic radiation; paraform; pressure; prototype; repository; sample fixation; seal; theories; therapeutic development

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.

描述（由申请人提供）：用福尔马林和石蜡固定组织样本是一种成熟的方法，用于组织的长期储存，以备以后的组织学分析。这些福尔马林固定石蜡包埋组织也可用于DNA， RNA或蛋白质的提取，以进行回顾性分子研究。不幸的是，传统的保存方法不能保证均匀固定，经常导致样品周围分子过度交联，深度降解缓慢。因此，抗原检索、基因组或蛋白质组学生物标志物鉴定研究可能产生不准确或误导性的结果。对其他固定方法的尝试，如对流加热或微波辐射治疗，也被证明是不充分的，因为它们不能产生均匀的产物，或导致生物分子谱的扰动。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标准化组织固定过程，并进行精确的热力学控制。高压已被证明可以促进甲醛快速均匀地渗透到组织块中。提出的组织保存系统将通过增强固定剂的渗透来更好地保存组织，同时保存组织结构。这种改进将导致从保存的样本中产生更好的数据，这可能导致更快的疾病诊断和治疗方法的成功开发。1