Develop extremely-high-yield (5-10x conventional methods) tissue microarray techn

描述(由申请人提供)：我们(阵列科学公司，LLC)提议对我们正在申请专利的组织微阵列(TMA)高产生产方法和仪器进行详细研究和开发。我们的TMA生产方法在利用标准组织学包埋介质(如石蜡)的液态方面是新颖的，通过这样做，我们实现了对宝贵生物组织资源的前所未有的利用水平。这项技术的成果在癌症诊断中被证明是非常有益的，它已经在一个主要的免疫组织化学(IHC)实验室产生了超过130万张TMA切片用于载玻片对照，在IHC熟练测试中，我们作为美国病理学家学会的供应商达到了新的产量水平。传统的TMA生产方法每TMA块生产100-300张幻灯片，而我们的方法通常每TMA块生产1500张幻灯片，大大减少了源组织的浪费水平。理论收益率仍然是我们目前达到的水平的两倍，有两个基本要素是重复实现最大收益率和实现我们方法的全部潜在好处所必需的。首先，需要对热流体力学和组织力学有更深入的了解，这是我们方法的基础，并决定了高产量TMA块在组织学上的成功。同时，根据这一改进的理解，需要对仪器进行设计更改，以处理与我们目前仪器中的流体和热流略有不同的情况。仪器中几个特定功能的自动化和计算机控制是这项工作的重要部分，并将成为第一阶段和第二阶段之间的技术桥梁。在拟议的研究中，将进行一项系统研究，以恢复一半至所有剩余可用产量，利用(A)用于非破坏性TMA块测试的MicroCT扫描(“虚拟组织学”)与自动图像分析相结合，将TMA切片缺陷与TMA块构建过程的人工制品相关联；(B)改进方法和仪器，以更好地控制两阶段块构建过程的流体和固体力学；(C)阵列科学程序的自动化，以提高技术的一致性和对新用户的可移植性。还将系统地研究阵列科学方法在组织来源制备方面的多功能性，从传统的(FFPE块取芯)方法，到液体悬浮液中高度碎裂的固体组织，再到细胞培养材料的使用。对MicroCT切片图像和相应组织切片的显微照片进行定量比较分析，以及组织学中的切片成品率测量，有望成为一种新的和强大的验证MicroCT作为一种非破坏性“虚拟组织学”的方法，以获得关于高产量TMA构建方法的敏感性和可控性的明确结论。

DESCRIPTION (provided by applicant): We (Array Science, LLC) propose to perform detailed research and development of our patent-pending method and instrument for high-yield production of tissue microarrays (TMAs). Our approach to TMA production is novel in leveraging the liquid phase of standard histologic embedding media such as paraffin, and in so doing we achieve unprecedented levels of utilization of precious biological tissue sources. The outcome of this technique has proven highly beneficial in cancer diagnostics, where it has produced over 1.3 million TMA sections for on-slide controls in a major immunohistochemistry (IHC) laboratory, and in IHC proficiency testing, where we have achieved new levels of yield as a supplier to the College of American Pathologists. Whereas conventional TMA production methods yield 100-300 slides per TMA block, our method routinely produces 1500 slides per TMA block, with highly reduced levels of waste of source tissue. Theoretical yield is still up to twice our currenty achieved level, with two essential elements necessary for repeatably achieving maximum yields and realizing the full potential benefits of our method. First, a deeper understanding is required of the thermal-fluid dynamics and tissue mechanics that underly our method and dictate the success of a high-yield TMA block in histology. Concurrently and in light of this improved understanding, design changes to the instrument are required that will handle fluid and heat flow somewhat differently than in our instruments to date. Automation and computer control of several specific functions in the instrument are an important part of the effort, and will serve asa technical bridge between Phase I and Phase II. In the proposed research, a systematic study will be performed to recover half to all of the remaining available yield, utilizing (a) microCT scanning for non-destructive TMA block testing ("virtual histology") combined with automated image analysis to correlate TMA section flaws with artifacts of the TMA block building process; (b) method and instrument refinements to better control the fluid and solid mechanics of the two-phase block building process; (c) automation of the Array Science process for improved uniformity and portability of the technology to new users. The versatility of the Array Science method will also be systematically studied as regards tissue source preparations, from conventional (FFPE block coring) methods, to highly fragmented solid tissue in liquid suspension, to the use of cell culture material. Quantitative comparative analyses of microCT slice images and micrographs of their corresponding histologic sections, and section yield measurements in histology, hold promise as a novel and powerful validation of microCT as a nondestructive "virtual histology" to achieve definitive conclusions about sensitivity and control of the high-yield TMA construction method.