Upgrade of Aquilos to Aquilos 2 cryo-focused ion beam scanning electron microscope. In our original proposed budget, we requested ~$1M for a focused ion beam instrument. We did not specify the manufacturer of such instrument (i.e. Thermo Fisher or Zeiss). The recent announcement from Thermo Fisher of the Aquilos 2 (~$1.38M), offers multiple exciting features. It has the capabilities of performing lift-out, slice and view and automated milling with 20+ lamellae in 13 hours of operation. These results suggest that cryoFIB can become an automated procedure on very thick, high-pressure vitrified samples. This would be a highly sought-after capability of our Service Center, because we anticipate that there will be a significant demand if we can begin to work with tissues tens to hundreds of microns in thickness. These performance specifications and the results shown by Thermo Fisher are exciting. If funded, we will also investigate whether Zeiss can offer a similar or superior instrument before we make a purchase decision. Certainly, we will consult with our European colleagues who have the prototypes of the new instrument in terms of its robustness as a user facility as in our Center.
Statement We propose to establish the Stanford-SLAC CryoET Specimen Preparation Service Center (SCSC) to meet the emerging national need for cryoET as a structural biology tool. The Center facility will enable external users to streamline the preparation of samples through providing access to, and support for, advanced cryo-specimen preparation techniques (cryoFLM, cryoFIB, and cryoEM/ET) for a wide range of samples with biomedical applications. The Center will offer a comprehensive program to train and cross-train scientists, regardless of their institutional affiliations, to become independent cryoET investigators.
