Advancing Multi-Color EM via Direct Detector-enabled 4D-STEM

项目总结/摘要 这一竞争性补充申请要求资金采购高速直接电子计数探测器 能量过滤透射电镜（EFTEM）这个探测器将使我们能够加速验证， 正在开发的新型多标记元素探针的性能基准测试，以扩大影响， 电子显微镜（ColorEM）。这项工作将在一个积极的背景下进行， 该研究项目正在推进ColorEM新方法的开发，采用新技术 用于超高速4D-STEM（5 R 01 GM 138780 -03）。该资助项目正在进行可行性研究 并克服了以下技术难题：1）用元素探针制作多标记电镜样品 针对ColorSTEM优化的组合（并可扩展到ColorEM的所有方法）; 2）首次使用时进行故障排除 潜在的变革性新技术，用于STEM检测和相关的原子数能力， (Z)对比成像;以及3）制定用于处理/分析非常大的原始数据集的方法 以便加速元素区分/信号解释并允许显示ColorSTEM结果。的 超快直接电子探测器的要求，将使我们能够加速和提高我们的记录能力， 比较和优化性能元素探头组合（信噪比和差分 对比度），并提供对我们的定量性能的更灵敏和准确的验证。 数据处理程序，与标记化学品携手工作，使我们能够可靠地 准确地将从一个选择性标记的结构记录的信号与另一个的特定信号区分开。 所需的设备项目是Apollo（Direct Electron，LP）、低噪音、高速电子计数、直接 检测设备（DDD）的透射电子显微镜，它具有行业领先的信号， 噪声和吞吐量规格，使我们能够更快速、更准确地验证 新的元素探测器，从而加快完成目标1和3的母奖。

PROJECT SUMMARY / ABSTRACT This competitive supplement application requests funds to procure a high speed direct electron counting detector for analytical Energy Filtered TEM (EFTEM). This detector will allow us to accelerate the validation and performance benchmarking of new multi-label, elemental probes under development to scale the impact and accessibility of multicolor electron microscopy (ColorEM). This work will be conducted in the context of an active research project that is advancing the development of a novel method for ColorEM, employing new technologies for ultra-high-speed 4D-STEM (5 R01GM138780-03). This funded project is performing the feasibility studies and surmounting the technical challenges to: 1) make multi-labelled EM specimens with elemental probe combinations optimized for ColorSTEM (and extensible to all methods for ColorEM); 2) troubleshoot the first use of potentially transformative new technologies for STEM detection and associated capabilities for atomic-number (Z) contrast imaging; and, 3) work out the methodology for processing/analysis of very large primary data sets so as to accelerate elemental differentiation/signal interpretation and allow for display of ColorSTEM results. The ultra-fast direct electron detector requested, will allow us to accelerate and sharpen our ability to record, compare, and optimize the performance elemental probe combinations (for both signal-to-noise and differential contrast) as well as provide for a more sensitive and accurate validation of the quantitative performance of our data processing routines, which work hand-in-hand with the labeling chemistry to allow us to reliably and accurately discern the signal recorded from one selectively labeled structure from the specific signal of another. The equipment item requested is an Apollo (Direct Electron, LP), low-noise, high-speed electron-counting, direct detection device (DDD) for transmission electron microscopy, which possesses an industry-leading signal-to- noise and throughput specification that will allow us to more rapidly and accurately validate the performance of new elemental probes, thus accelerating the completion of Aims 1 and 3 of the parent award.