Training to use the Sphera is fairly intensive. It is expected that the user will be able to do more than just push the buttons.
Proficiency with the instrument without direct supervision is expected. If the basics are not learned and mastered the student still may be able to get images but they won't be of research quality. Training averages about 15 – 20 hours per
individual and it is usually scheduled in half day a week sessions. During most of that time the student will be using a supplied standard instead of their own samples. Accommodations in the training routine are made
for those individuals who have had prior microscopy experience. Users are given a microscope account on the Sphera,
access to the on-line Scheduler
, access to the UCSD
CryoEM list serve, and an account on other computers in the facility for the temporary storage of digital data. Sessions
are broadly based around the following topics:
- Initial meeting to describe the process and find out the research needs of the student – 1/2 hour
- Description of the column, control pads and the vacuum system – 3 hours
- Description of the Tecnai operating system, personal customization of the Tecnai user account, and sample holder insertion and removal – 3 hours
- Microscope log-on procedures, finding the beam, condenser aperture alignment, condenser astigmatism correction, gun alignments and other associated alignments on that page, eucentric height correction, objective aperture alignment, and objective lens rotation center alignment. Practice on insertion of the sample holder, supervised microscope practice time and microscope shutdown. – 3 hours
- Introduction to critical objective lens focusing including an introduction to the principles of scattering and phase contrast. Practice with procedures in step 4. Additional microscope practice time – 3 hours
- Introduction to objective lens astigmatism correction. Practice with procedures in steps 4 and 5. Additional microscope practice time – 3 hours
- Additional instruction in Digital Micrograph, file saving and file transfer and practice time on all procedures – 2 hours
- Future sessions with the Facility Manager will depend on the level of competence of the student and the need for further close supervision. At this time the student is encouraged to bring in his/her own samples.
- When proficiency is demonstrated, the student schedules time for themselves on the Scheduler during the normal working hours. At this time it is not necessary for the Facility Manager to be present. Twenty-four hour access to the facility may be granted to students who have shown an expertise on the instruments. This may occur after the student has had an extended period of time of independent use without encountering any problems.
Additional training sessions may be scheduled based on the needs of the user. Such sessions could include:
- Data collection on film and use of the darkroom
- Low dose training
- Preparation of continuous carbon and holey carbon support films
- Negative staining
- Advanced microscope alignment
- Cryo-microscopy (multiple sessions)