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

描述（由申请人提供）：我们（Array Science， LLC）提议对我们正在申请专利的用于组织微阵列（tma）高产生产的方法和仪器进行详细的研究和开发。我们的TMA生产方法在利用标准组织包埋介质（如石蜡）的液相方面是新颖的，这样做我们实现了对珍贵生物组织来源的前所未有的利用水平。这项技术的成果已被证明在癌症诊断方面非常有益，它已在一个主要的免疫组织化学（IHC）实验室生产了超过130万张TMA切片用于载玻片对照，在免疫组织化学水平测试中，作为美国病理学家学院的供应商，我们的产量达到了新的水平。传统的TMA生产方法每个TMA块生产100-300片，而我们的方法通常每个TMA块生产1500片，大大减少了源组织的浪费。理论产量仍然是我们目前达到的水平的两倍，有两个必要的因素可以重复实现最大产量，并实现我们方法的全部潜在效益。首先，需要对热流体动力学和组织力学有更深入的了解，这是我们方法的基础，也是高产量TMA块在组织学上取得成功的关键。同时，鉴于这种改进的理解，需要对仪器的设计进行更改，以处理流体和热流，与我们迄今为止的仪器有所不同。仪器中几个特定功能的自动化和计算机控制是工作的重要组成部分，并将作为第一阶段和第二阶段之间的技术桥梁。在拟议的研究中，将进行一项系统的研究，以恢复一半到所有剩余的可用产量，利用(a)微ct扫描进行非破坏性TMA块测试（“虚拟组织学”），结合自动图像分析，将TMA切片缺陷与TMA块构建过程中的伪影联系起来；(b)方法和仪器的改进，以更好地控制两相砌块构建过程的流体和固体力学；(c)阵列科学过程的自动化，以提高技术对新用户的统一性和可移植性。Array Science方法的多功能性也将在组织源制备方面进行系统研究，从传统的（FFPE块取芯）方法，到液体悬浮液中高度碎片化的固体组织，再到细胞培养材料的使用。通过对微ct切片图像和相应组织切片的显微照片的定量比较分析，以及组织学中切片产量的测量，有望对微ct作为一种无损的“虚拟组织学”进行新颖而有力的验证，从而得出关于高产量TMA构建方法的灵敏度和控制的明确结论。

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.