Modern Cellular and Molecular Techniques to Study the Human Temporal Bone

研究人类颞骨的现代细胞和分子技术

• 批准号: 10604912
• 负责人: Akira Ishiyama
• 金额: $ 66.72万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-19 至 2027-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10604912
• 关键词: 3-Dimensional; Anatomy; Animal Model; Animals; Archives; Autopsy; Biological; Bone Tissue; Celloidin; Clinical; Collaborations; Collection; DNA; DNA Fragmentation; DNA Sequence; DNA sequencing; Digital Libraries; Ear; Genes; Harvest; Hematoxylin and Eosin Staining Method; High Resolution Computed Tomography; Histopathology; Human; Image; Imaging Techniques; Immunofluorescence Immunologic; In Situ Hybridization; Laboratories; Labyrinth; Libraries; Magnetic Resonance Imaging; Messenger RNA; Methods; Modality; Modernization; Molecular; Molecular Biology Techniques; Morphology; Mus; National Institute on Deafness and Other Communication Disorders; Neurons; Nucleic Acids; Organ; Phase; Process; Proteins; Protocols documentation; RNA; Registries; Research; Research Personnel; Resolution; Resources; Scientist; Services; Specimen; Stains; Structure; Techniques; Temporal bone structure; Testing; Time; Tissue Banks; Tissue Fixation; Tissues; Update; Visualization; X-Ray Computed Tomography; bone imaging; contrast imaging; deep learning; design; digital; experience; hands on instruction; high resolution imaging; human imaging; image archival system; imaging modality; improved; interest; meetings; novel; preservation; prospective; reconstruction; sample fixation; spiral ganglion; symposium; synchrotron radiation; three-dimensional visualization; tissue processing; tool; ultra high resolution; virtual; web site

PROJECT SUMMARY In the present proposal we will investigate new tissue processing protocols designed to preserve DNA, as well as the morphology. We will preserve, process, and make available human ear tissue to inner ear basic researchers they will be able to compare with their animal models. We have experience using high-resolution advanced imaging techniques to the postmortem temporal bones, and also the membranous structures of the inner ear. We propose to develop and apply these imaging techniques systematically and to correlate the imaging findings with the temporal bone histopathology aiming to visualize structures with non-destructive techniques. In Specific Aim 1) we will serve as part of a collaborative network providing technical services for procuring, preparing, sectioning, and distributing high-quality human inner ear tissues aiming to increase interest in the use of human inner ear tissues for basic scientists. We will continue to procure post- mortem temporal bones aiming to minimize post-mortem times and will facilitate and reach out to additional prospective researchers through the collaborative network and through meetings and provide human inner ear tissues of high-quality. We will create hematoxylin and eosin digital shareable library with our archive of temporal bones, and this library will be shared through the National Temporal Bone Registry to scientists. We will disseminate protocols for temporal bone collection, processing, and inner ear tissue processing and provide hands-on instruction on these protocols on demand and via zoom conferences. In Specific Aim 2, we will develop new tissue collection and processing techniques for temporal bones by using animal temporal bones to develop techniques which preserve DNA, mRNA and proteins and preserve morphology. We aim to develop new tissue processing protocols which will allow for the use of immunofluorescence, DNA extraction and sequencing, and the application of RNAScope, a powerful in situ hybridization tool. A new protocol will be developed to further optimize sequencing the DNA from the extracted DNA using temporal bones specimens to correlate the gene sequencing with the histopathology. In Aim 3. We will improve temporal bone inner ear visualization using novel non-destructive imaging techniques which we will correlate with histopathology. In Aim 4, we will develop a deep learning-based stain transformation framework to visualize specific anatomical structures such as the spiral ganglion neurons to obtain accurate histopathological analysis.

项目概要 在本提案中，我们还将研究旨在保存 DNA 的新组织处理方案 作为形态。我们将保存、处理人耳组织并将其提供给内耳基础 研究人员将能够与他们的动物模型进行比较。我们有使用高分辨率的经验 先进的成像技术可用于死后颞骨以及颞骨的膜结构 内耳。我们建议系统地开发和应用这些成像技术，并将 颞骨组织病理学成像结果旨在以非破坏性方式可视化结构 技术。在具体目标 1) 中，我们将作为协作网络的一部分，为以下领域提供技术服务： 采购、准备、切片和分发高质量的人体内耳组织，旨在增加 基础科学家对使用人类内耳组织感兴趣。我们将继续采购尸检 颞骨旨在最大限度地减少尸检时间，并将促进和接触更多的潜在人员 研究人员通过协作网络和会议提供了人类内耳组织 高质量。我们将利用我们的颞骨档案创建苏木精和伊红数字共享图书馆， 该库将通过国家颞骨登记处与科学家共享。我们将传播 颞骨采集、处理和内耳组织处理的协议，并提供实践操作 按需并通过 Zoom 会议提供有关这些协议的说明。在具体目标 2 中，我们将开发新的 利用动物颞骨开发颞骨组织采集及处理技术 保存 DNA、mRNA 和蛋白质并保留形态的技术。我们的目标是开发新的组织 允许使用免疫荧光、DNA 提取和测序的处理方案，以及 RNAScope（一种强大的原位杂交工具）的应用。将制定新协议以进一步 使用颞骨样本对提取的 DNA 进行优化测序，以关联基因 与组织病理学测序。在目标 3 中，我们将使用以下方法改善颞骨内耳可视化 新颖的无损成像技术，我们将其与组织病理学相关联。在目标 4 中，我们将开发 基于深度学习的染色转换框架，用于可视化特定的解剖结构，例如 螺旋神经节神经元以获得准确的组织病理学分析。