EAGER: Live Cell Imaging using Low Dose 4DSTEM

现代生物学研究的驱动力是需要以更精细的细节水平观察活细胞和结构。 大多数活细胞成像工作依赖于可见光或荧光成像，因为更高分辨率的方法，如透射电子显微镜（TEM）使用更高的能量和对细胞致命的样品制备方法。 因此，目前束流对活细胞的损伤的限制阻止了该领域进入纳米级活生物成像的下一阶段。 该奖项支持的研究人员将通过开发新的非致命TEM成像方法来克服这一挑战，以实现无与伦比的细节活细胞成像。 从这些研究中获得的知识将被整合到UT奥斯汀的“实用电子显微镜”课程中。此外，本科生研究项目将在UT奥斯汀提供作为尤里卡计划的一部分，并积极鼓励代表性不足的群体参与。这些研究还将在公共科学活动中展出，如UT Austin的年度“女孩日”，超过8，000名小学和中学生参观UT Austin并获得科学和工程方面的实践经验;以及在黑人历史月期间UT Austin的年度“黑人卓越STEM”活动。研究人员还将参加一年一度的“探索UT”，这是一个开放日，成千上万的外地居民参观校园，参加高接触，互动，动手程序。活细胞和生物过程的纳米级成像将使用先进的低剂量4D扫描透射电子显微镜（4D-STEM）进行。将使用液体细胞原位保持器和高灵敏度超快像素化直接电子检测器进行高对比度成像，以观察具有非致死电子剂量的活细胞。研究将从观察细胞膜与纳米级物体（如外来体和病毒）的相互作用开始，以提供各种生物过程中活细胞的新动力学见解。以纳米级的分辨率观察活细胞开辟了一个新的领域，即生物学中的真实的时间动力学，其长度尺度是目前无法达到的。通过该奖项开发的成像方法，信息学和实验程序将用于广泛的生物系统研究，包括细胞间通讯，病毒：细胞膜相互作用，细菌复制和细胞运动。 这些研究的结果将发表在同行评审的期刊上，在科学会议上提出，并通过公共宣传活动。该奖项由生物基础设施部和分子与细胞生物科学部共同支持。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Modern biological research is driven by a need to observe living cells and structures at ever-finer levels of detail. Most of the live-cell cell imaging work relies on visible or fluorescent light imaging because higher resolutions methods, like transmission electron microscopy (TEM) use higher energies and sample preparation methods that are lethal to cells. The current limitation of beam induced damage to live cells is thus preventing this field from moving to the next phase of nanoscale live biological imaging. Researcher supported by this award will overcome this challenge by developing new, non-lethal TEM imaging methods to enable live cell imaging in unparalleled detail. Knowledge gained form these studies will be integrated into the ‘Practical Electron Microscopy’ course at UT Austin. In addition, undergraduate research projects will be offered at UT Austin as part of the EUREKA program and participation from underrepresented groups will be actively encouraged. The studies will also be featured at public science activities like the annual ‘Girl Day’ at UT Austin, where over 8,000 elementary and middle school students visit UT Austin and have hands-on experience with science and engineering; and at the annual ‘Black Excellence in STEM’ event at UT Austin during Black History Month. The researchers will also participate in the annual ‘Explore UT’, the open day where thousands of out-of-town residents visit campus to participate in high-touch, interactive, hands-on programs. Nanoscale imaging of live cells and biological processes will be undertaken using advanced low-dose 4D scanning transmission electron microscopy (4D-STEM). High contrast imaging to observe live cells with non-lethal electron doses will be done using a liquid cell in situ holder and highly sensitive ultrafast pixelated direct electron detectors. Studies will start with the observation of cell membranes interacting with nanoscale objects such as exosomes and virus to provide new dynamical insights of live cells in various biological processes. Observing live cells with nanoscale resolution opens a new field of real time dynamics in biology at length scales that are current inaccessible. The imaging methods, informatics and experimental procedures developed through this award will be used in studies for a wide range of biological systems, including intercellular communication, virus:cell membrane interactions, bacteria replication and cell motility. Results from the studies will be published in peer-reviewed journals, presented at scientific meetings, and through public outreach events. This award is jointly supported by the Division of Biological Infrastructure and the Division of Molecular and Cellular Biosciences.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.