This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. ABSTRACT: In the reporting period of 4/1/05-3/31/06, the F30 microscope has been used to generate an extraordinary number of tilt-series (1107), despite a number of serious service problems and the installation of the Gatan Ultracam at the back of the GIF. Together these caused 418 hours (10.5 weeks) of down time. Due to the installation of the Gatan Ultracam and upgrades to our Ultrascan camera that is in front of the GIF, more time than usual has been spent on development of various software and hardware components (72.5 hours). The Ultrascan now has 4 readout ports, which has reduced the time it takes to acquire a tilt series from 45 min to about 20 min. If you take into account the service downtime, the F30 was in use 46 hours/week (1904 actual hours at the microscope for 41.5 weeks). Figure 1 illustrates the F30 usage based on the total number of hours used (2740.25hrs/52weeks). Training was limited to supervised, productive sessions with students who had previously been trained on the Tecnai F20 microscope, so the time is incorporated in either cryo or plastic tomography as appropriate. Figure 2 shows the division of different types of tomography work conducted on the F30 during the reporting period: single or dual-axis tomography of plastic-embedded sections at room temperature (Plastic TS), cryo tomography (Cryo TS), or montage tomography (Montage TS).
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