Developing a device for visualization of large-scale cryopreservation

开发大规模冷冻可视化装置

• 批准号: 8246407
• 负责人: YOED RABIN
• 金额: $ 11.65万
• 依托单位: CARNEGIE-MELLON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8246407
• 关键词: Address; Automobile Driving; Basic Science; Biocompatible Materials; Biological Preservation; Cells; Computer software; Contracts; Coupled; Cryopreservation; Cryoprotective Agents; Crystallization; Data; Development; Device or Instrument Development; Devices; Equilibrium; Event; Evolution; Fracture; Glass; Health; High temperature of physical object; Ice; Image; Imagery; Kinetics; Measurement; Mechanical Stress; Mechanics; Methodology; Methods; Modeling; Monitor; Organ; Outcome; Phase; Plastics; Prevention; Process; Production; Protocols documentation; Recovery; Research Proposals; Rewarming; Sampling; Scanning; Seeds; Solid; Solutions; Specimen; Staging; Stress; Structure; Surface; System; Techniques; Temperature; Testing; Time; Tissue Model; Tissues; Toxic effect; Vial device; Video Recording; Viscosity; Work; cold temperature; cryobiology; design; innovation; meetings; movie; polarized light; prevent; quality assurance; research and development; software development; success; tissue/organ preservation; tool; transplantation medicine; vitreous state

DESCRIPTION (provided by applicant): Preservation of tissues and organs at very low temperatures-cryopreservation-appears to be only feasible alternative to providing high quality biomaterial for transplantation medicine. While techniques for cryopreservation of single cells and small tissue structures are well established, cryopreservation techniques for bulky tissues and organs are still at the developmental stage. One of the most promising techniques for large-scale cryopreservation is known as vitrification, where ice crystallization is suppressed, and the biological material is trapped in a glassy-like state (vitreous in Latin means glassy). Vitrification is achieved by introducing cryoprotective agents (CPAs) into the tissue, which prevent the formation of ice crystals when cooled rapidly enough. While ice prevention is the key for successful cryopreservation by vitrification, the high cooling rate required for vitrification drives another potentially devastating effect, which is thermo-mechanical stress (driven by the tendency of the material to contract with the decreasing temperature), with fracture formation as its most dramatic outcome. The outcome of any cryopreservation process cannot merely be evaluated at the end points-the storage and room temperatures-where evidence of physical events that took place during the cooling or rewarming stages may have disappeared by the time that the specimen has reached equilibrium. A means to continuously monitor the process of cryopreservation and document such evidence represents an unmet need. This research proposal concerns the development of a new device for visualization of physical effects associated with crystallization, vitrification, and fracture formation during cryopreservation of large specimens. The innovation in the conceptual design of the new device is that it will be integrated-as an add-on unit-with commercially used and widely available freezer for cryobiology applications. Three principal usages are envisioned for the proposed device: (i) as a basic research tool, to explore physical effects associated with cryopreservation; (ii) as a research and development tool, to develop new cryopreservation protocols; and (iii) as a quality assurance tool, to be incorporated with mass production of cryopreserved products. Four specific aims have been set for the development of the scanning cryomacroscope: (1) to develop a scanning device for a specimen placed in the chamber of an available controlled-rate freezer; (2) to develop software for integration of video recording, registration, and temperature data; (3) to demonstrate the application of the integrated system on crystallization and vitrification processes; and, (4) to integrate polarized light into the scanning device and to develop related methodology. The application of polarized light has been proven to enhance identification of ice nucleation, as well as a means to investigate the development of mechanical stress in glasses and other solid transparent media.

描述（由申请人提供）：在极低温度下保存组织和器官-冷冻保存-似乎是为移植医学提供高质量生物材料的唯一可行替代方案。虽然单细胞和小组织结构的冷冻保存技术已经很成熟，但大体积组织和器官的冷冻保存技术仍处于开发阶段。大规模冷冻保存最有前途的技术之一是玻璃化，其中冰结晶被抑制，生物材料被困在玻璃状状态（拉丁语中的玻璃体是玻璃状的意思）。玻璃化是通过向组织中引入低温保护剂（CPA）来实现的，当足够迅速地冷却时，可以防止冰晶的形成。虽然防冰是通过玻璃化成功冷冻保存的关键，但玻璃化所需的高冷却速率驱动了另一种潜在的破坏性影响，即热机械应力（由材料随着温度降低而收缩的趋势驱动），其中裂缝形成是其最引人注目的结果。任何低温保存过程的结果不能仅仅在终点（储存温度和室温）进行评估，因为在冷却或复温阶段发生的物理事件的证据可能在标本达到平衡时消失。持续监测冷冻保存过程并记录此类证据的手段代表了未满足的需求。本研究建议涉及一种新的设备的发展与结晶，玻璃化，和大标本的冷冻保存过程中的骨折形成的物理效应的可视化。新设备概念设计的创新之处在于，它将作为一个附加单元与商业上广泛使用的冷冻机集成，用于低温生物学应用。预期所提出的装置有三种主要用途：（i）作为基础研究工具，探索与冷冻保存相关的物理效应;（ii）作为研究和开发工具，开发新的冷冻保存方案;以及（iii）作为质量保证工具，与冷冻保存产品的大规模生产相结合。扫描低温显微镜的发展有四个具体目标：（1）研制一种用于在现有的可控速率冷冻室中放置样品的扫描装置;（2）研制一种用于集成视频记录、记录和温度数据的软件;（3）演示集成系统在结晶和玻璃化过程中的应用;以及（4）将偏振光集成到扫描装置中并开发相关的方法。偏振光的应用已被证明可以增强冰成核的识别，以及研究玻璃和其他固体透明介质中机械应力发展的手段。