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.

项目摘要 在本提案中，我们将研究旨在保留DNA的新组织加工方案 作为形态。我们将保存，处理并使人耳组织可提供内耳的基础 研究人员将能够与动物模型进行比较。我们有使用高分辨率的经验 高级成像技术到验尸颞骨，以及膜结构的膜结构 内耳。我们建议系统地开发和应用这些成像技术，并将 颞骨组织病理学的成像发现旨在可视化无损的结构 技术。在特定目标中1）我们将作为一个协作网络的一部分，为技术服务提供技术服务 采购，准备，分割和分发旨在增加的高质量的人内耳组织 对使用人内耳组织的兴趣作为基础科学家。我们将继续采购验尸 临时骨骼旨在最大程度地减少验尸时代，并将促进并与其他潜在的潜在 研究人员通过协作网络和会议，并提供人类的内耳组织 高质量。我们将使用我们的颞骨档案创建苏木精和曙红数字可共享图书馆， 该图书馆将通过国家颞骨注册表与科学家共享。我们将传播 颞骨收集，加工和内耳组织加工的协议并提供动手 按需和缩放会议的这些协议的指示。在特定目标2中，我们将开发新的 通过使用动物颞骨发育的颞骨收集和加工技术 保留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 处理方案将允许使用免疫荧光，DNA提取和测序以及 Rnascope的应用，一种强大的原位杂交工具。将开发一个新协议以进一步 使用颞骨标本优化从提取的DNA的测序DNA以相关 与组织病理学进行测序。在AIM 3中。我们将使用 新型的非破坏性成像技术，我们将与组织病理学相关。在AIM 4中，我们将发展 一个基于深度学习的染色转化框架，可视化特定的解剖结构，例如 螺旋神经神经元获得准确的组织病理学分析。