Improved Imaging for Precise Crytpic Anomaly Detection

描述(申请人提供)：该项目将进一步发展 自动检测工具和图像分析技术，以检测 染色体重排，这是常规方法难以检测到的 细胞遗传学。它结合了两个创新的FISH探针：“亚端粒”(STFISH) 和“多重”(MFISH)，具有复杂的图像分析，用于放映 患者发育性残疾。 在第一阶段项目中，我们评估了(1)实施的可行性 染色体中期的自动识别，以及(2)使用复杂的 图像分析，用于准确检测亚微观重排。我们也 比较了两种FISH技术在密码学上的效率和精度 重排检测。我们用STFISH分析了8个样品，6个样品 使用MFISH。我们的实验结果表明，使用STFISH探针 大小从300Kbp到30Mbp，允许描绘染色体 有8/10的样品发生了重排。其余2个样本被考虑 半隐晦的，因为亚端粒探针集不包含染色体 22号染色体p臂的特异探针。因此，来自中国的卫星 端粒探针组不能识别22P。MFISH检测结果为 有效检测3/6例染色体重排，其中2例为 半隐蔽的，其中一个有3向重排，包括5Mbp的插入。我们 为自动化系统开发的软件组件，包括自动对焦、 24井载玻片扫描(Cytocell TM)，中期发现。最后， 我们估计了通过添加改进的 两种成像方法均以FISH技术为主。 在第二阶段，我们将进一步开发这项技术，并在临床上进行测试 环境。具体地说，对于MFISH，我们将优化软件以增强 在检测重排方面的性能。对于STFISH，我们将(1)确定 自动化系统的扫描和中期发现率，(2)测试 临床环境中的原型；以及(3)评估商业 该仪器的可行性。然后，第三阶段将使这项新技术 可用于商业化。

DESCRIPTION (provided by applicant): This project will further develop automated instrumentation and image analysis techniques to detect cryptic chromosomal rearrangements, which are difficult to detect by conventional cytogenetics. It combines two innovative FISH probes: "SubTelomeric" (STFISH) and "Multiplex" (MFISH), with sophisticated image analysis, for screening patients developmental disabilities. During the Phase I project, we evaluated the feasibility of (1) performing automated identification of chromosome metaphases, and (2) using sophisticated image analysis for accurate detection of submicroscopic rearrangements. We also compared the efficiency and precision of the two FISH techniques for cryptic rearrangement detection. We analyzed 8 samples using STFISH, and 6 samples using MFISH. Our experimental results show that, the use of STFISH probes ranging in size from 300Kbp to 30Mbp, permitted the delineation of chromosomal rearrangements in 8/10 samples. The remaining 2 samples were considered half-cryptic, since the Subtelomeric probe set does not contain a chromosome specific probe for the p-arm of chromosome 22. Therefore, the satellites from 22p could not be identified with the telomere probe set. The MFISH assay was effective in detecting chromosomal rearrangements in 3/6 cases of which 2 were half-cryptic, and one had a 3-way rearrangement, including a 5Mbp insertion. We developed software components for an automated system, including auto focusing, slide scanning for 24 well slides (CytocellTM), and metaphase finding. Finally, we estimated the cost and time savings achievable, by the addition of improved imaging methods to both the FISH techniques. In Phase II, we will further develop the technology and test it in a clinical environment. Specifically, for MFISH we will optimize the software for enhanced performance in detecting rearrangements. For STFISH, we will (1) determine the scanning and metaphase finding rates of the automated system, (2) test the prototype in a clinical environment; and (3) evaluate the commercial feasibility of the instrument. Phase 3 will then make this new technology available for commercialization.