Energy filter with fast-counting direct electron detector

• Subject Area: Basic Research in Biology and Medicine
• Term: Funded in 2023
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 525040890

Researchers at the Julius-Maximilians-Universität Würzburg investigate the structural determinants by which biological assemblies support a diverse set of functionalities. In 2018 they established an electron cryo microscopy facility for high-resolution structure determination. This facility is used by researchers in and beyond Würzburg. At the heart of the platform is a 300 kV electron microscope with a Schottky type XFEG emitter and a direct electron detector. The detector enables data acquisition in linear mode and in counting mode. The latter gives a detective quantum efficiency, which is close to the theoretical limit. However, due to its low frame rate it limits the throughput to only about 700 movies per day. In contrast, linear mode allows data acquisition of up to 7,000 movies a day but with only about half of the detective quantum efficiency of the counting mode. Sparked by the success with data obtained at the facility, the ambitions of the researchers at the JMU have shifted towards more challenging projects such as smaller, and more dynamic complexes and ligand localization for drug discovery. The success of these projects depends on large data sets with some 5000 movies obtained in counting-mode. With the current equipment, acquisition of such data sets can take a week and more. Even worse, for smaller dynamic complexes (< 150 kDa), the scouting data obtained within 24 hours are often insufficient for judging whether a structure would likely be solved if a larger data set were obtained. Therefore, the current detection system with a slowly counting camera does not fulfill the needs and ambitions of the researchers at the JMU with respect to quality and quantity of image data obtained within a 24-hour slot. To establish whether a different detection system will be more suitable for the needs, researchers compared data obtained for the same sample in Würzburg and elsewhere. These comparisons showed that in addition to counting, energy filtration and zero-loss imaging are other important assets which enable more robust classification of small dynamic complexes and drastically improve the overall resolution. To bring the platform to the current leading standards in image acquisition, we apply for replacing the current detection system with an energy filter and a direct electron detector with fast counting capabilities.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Energiefilter mit schnell zählendem direkten Elektronendetektor
• Instrumentation Group: 5140 Hilfsgeräte und Zubehör für Elektronenmikroskope
• Applicant Institution: Julius-Maximilians-Universität Würzburg
• Leader: Professorin Dr. Bettina Böttcher