Biological and Cryogenic Transmission Electron Microscopy
Grant Jensen, PI
Alasdair McDowall, Director
The Beckman Resource Center for Transmission Electron Microscopy (BRCem) provides advice, training and access to high-end instrumentation that Caltech researchers need to perform state-of-the-art biological and/or cryogenic TEM, and seeks to further develop these technologies to better meet research objectives. The BRCem is located in the Jensen laboratory, which uses state-of-the-art electron cryo-microscopy techniques to understand the structure and function of large protein machines and their arrangement within cells.
The facility is equipped with two transmission electron microscopes for high-resolution imaging of cryogenic samples.
- F30: Tecnai Field Emission (FEG) 300keV TEM Polara (F30) equipped with a Gatan K2 Summit Direct Electron Detector and Gatan Energy Filter, the "Leginon" automated data collection software suite, as well as the UCSF Tomography automated tomography software.
- T12: Tecnai T12 LaB6 120keV TEM equipped with a Gatan Ultrascan 2k X 2k CCD, the "Leginon" automated data collection software suite, as well as the Serial EM automated software.
Access to the two electron microscopes is available for trained BRCem staff and collaborators as follows:
- F30: approximately 9 hrs/week during primetime (8 a.m. - 5 p.m.)
- T12: approximately 18 hrs/week during primetime (8 a.m. - 5 p.m.)
- Non-primetime hours are available to experienced and approved collaborators.
There are multiple options for collaboration, which will be discussed with BRCem staff:
- As a one-time shot, the BRCem staff will perform the work at a $90/hr service cost to the client. This is ideal, for example, for a short pilot feasibility study, or for the production of a final image for an existing well-characterized study.
- As a self-service project, the client will receive, with close supervision, the training required to be self-sufficient, in addition to protocols and basic consumables, at a cost of $90/hr. Thereafter, the client will continue the project independently and an access service charge of $50/hr will apply. Training may be organized in small group sessions.
- As a "co-author" collaboration, clients will work together with members of the Jensen group to complete the project. Each such project will be evaluated and budgeted on a case-by-case basis.
- As a total cost recovery project, the BRCem will provide a quote and time frame for the project, which may be outsourced to a third party. All costs are to be covered by the client.
(Statement Regarding "Fee for Service Charges" for BI Resource Electron Microscopy:
The internal Caltech rates of $90/hr and $50/hr for assisted and unassisted electron microscope use are calculated from the operational costs of instrument service contracts, consumables and associated salaries, relative to the user hours averaged over a three year period. These rates are comparable to surveyed nationwide electron microscopy service rates.)
Additional Equipment for Sample Preparation
- FEI Vitrobot vitrification device for plunge-freezing of aqueous single particles or thin cells
- BalTec HPM010 high-pressure freezer system for bulk tissue and cell vitrification.
- Leica AFS automated freeze substitution processor.
- Cryo and conventional ultramicrotome (Leica EM UC6/FC6).
- Nikon 90i computer-controlled epi-fluorescence microscope with Metamorph automation and imaging software.
- Ancillary units for carbon coating/rotation shadowing/plasma ionization.
The central laboratory area provides bench space for visitors, with temporary 4°C, -80°C and liquid nitrogen sample storage. Stocks of routine sample preparation chemicals are available, as are fume hoods and a Biohazard hood. Isolated rooms for light microscopy and ultramicrotomy procedures are part of the facility. High-capacity storage devices are available for loan to transfer image data.
For further information, visit http://www.jensenlab.caltech.edu
Personnel associated with the Center
||Grant J. Jensen
||Pamela J. Bjorkman
||Alasdair W. McDowall
||Mark S. Ladinsky
| Dr. Alasdair McDowall, Director
Broad Center - Room 20
Mail Code 114-96
Tel: (626) 395-8829
Mob: (626) 394-3467
Fax: (626) 395-8033