CryoElectron Microscopy Facility Description
(edited November 14, 2007)
The CryoElectron Microscopy Facility at UCSD is housed in the first level of
Bonner Hall on the Revelle College campus. The suite occupies
approximately 1700 sq. ft. and there is an additional 190 sq. ft. of wet lab
space (contains a fume hood, refrigerator, light microscopes, sonicators,
balance, etc) shared with another faculty member in the Division of Biology.
The CryoEM suite in Bonner Hall has nine rooms, including:
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Three microscope rooms designed to be optimal for cryoEM
experiments. Two of these rooms are designed to contain cryomicroscopes equipped with field-emission guns. The floors, walls and
ceiling surfaces are moisture sealed so that strict temperature and
humidity requirements can be maintained. Air supply into the room is
from fabric DuctSox™, air-dispersion tubes to reduce air currents in
the room. Sound dampening curtains surround the microscopes.
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A darkroom for developing photographic film plates with a nitrogenburst developing system
An equipment room that houses the microscope electronics and water
chillers
An office for the facility manager
A computer room to be used for initial processing of recorded image
data
A room that houses the dedicated Kathobar HVAC unit
A walk-in cooler and ante-room. The cooler is used to prepare vitrified
biological specimens. Such conditions are required to prevent
contamination of the cryogens with water vapor. The ante-room
houses some of the ancillary microscope equipment.
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The facility currently houses two FEI/Philips transmission cryo-electron
microscopes, including a 200kV Tecnai G2 Sphera, equipped with a LaB6
electron gun, and a 300KV Tecnai G2 Polara, equipped with a field emission
gun. The Polara has a modern stage design that allows examination of
vitrified samples at either liquid helium or liquid nitrogen temperatures. The
Sphera uses standard cryo-transfer stages developed by Gatan, Inc., two of
which are available for use by microscopists. Both microscopes are
completely computer controlled with the Tecnai GUI interface that runs
under the Microsoft Windows 2000 operating system. Both microscopes are
also equipped with tomography image acquisition and tomographic
reconstruction software. In addition, the Leginon software package,
developed by colleagues at the National Resource for Automated Molecular
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Microscopy in the Scripps Research Institute, is installed on both
microscopes.
Features of the FEI Tecnai G2 Sphera:
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Operates at voltages up to 200kV.
Cut-film holder for up to 56 films
Room-temperature, 70 degree tilt, sample holder
Gatan Ultrascan 1000 UHS CCD camera designed for a 200kV electron
source. The camera has a 4 megapixel (2k X 2k), Peltier-cooled, CCD
chip and is equipped with an ultra-high sensitivity phosphor
scintillator. The camera is mounted on-axis under the microscope
column and is connected via an optical cable to the microscope
computer.
Gatan 676 on-axis, intensified TV camera with a YAG scintillator and a
12 in, monochrome monitor.
Cryo package: includes an improved anticontaminator, Low-Dose
software, and cryo-cycle protection for the ion getter pump
Gatan Digital Micrograph software: operates the CCD camera and
permits various levels of analysis of recorded images
Two, NEC 20” LCD color monitors
Gatan HREM Autotune software: automatically sets image defocus and
stigmates CCD images
FEI Xplore3D tomographic software
SerialEM tomographic software (D. Mastronarde, University of
Colorado at Boulder)
Tecnai Scripting and Photomontage software
NRAMM Leginon automated data acquisition software
SerialEM tomographic software (D. Mastronarde, University of
Colorado at Boulder)
Two Gatan, 70-degree tilt cryo-transfer holders and one cryo-transfer
station
Gatan Dry Pumping Station: used to pump out cryo-holders for
improved performance
Miscellaneous ancillary equipment: e.g. a Haskris water circulator;
ZEM water circulator; film desiccator; etc.
Features of the FEI Tecnai G2 Polara
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Electron gun is a Schottky field emitter.
Operates at voltages up to 300kV.
Multi-specimen transfer device: has 6 storage positions for cartridges
that hold 3 mm EM grids
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Cryoworkstation: for mounting EM grids into the cartridges and to load
the cartridges into the multi-specimen transfer device. The
workstation has a loading area, a vacuum system and an airlock for
docking to the transfer device. The workstation can be rolled out of
the way when not needed.
Improved vacuum system: consists of a rotary pump and buffer tank,
an oil diffusion pump, a turbo-molecular pump, and four ion getter
pumps
Computer-controlled, ultra-stable goniometer that can be cooled down
to either liquid nitrogen or helium temperatures with the ability to tilt
up to 70 degrees.
Cut-film holder for up to 56 films
Gatan Ultrascan 4000 UHS CCD camera designed for a 300kV electron
source. The camera has a 16 megapixel (4k X 4k), Peltier-cooled,
CCD chip and is equipped with an ultra-high sensitivity phosphor
scintillator. The camera is mounted on-axis under the microscope
column and is connected via an optical cable to the computer of the
microscope.
Cryo package: includes an improved anticontaminator “cryobox”, LowDose software, and cryo-cycle protection for the ion getter pumps
Gatan Digital Micrograph software: operates the CCD camera and
permits various levels of analysis of recorded images
Two, NEC 20” LCD color monitors
Gatan HREM Autotune software: automatically sets image defocus and
stigmates CCD images
FEI Xplore3D tomographic software
SerialEM tomographic software (D. Mastronarde, University of
Colorado at Boulder)
Tecnai Scripting and Photomontage software
NRAMM Leginon automated data acquisition software
SerialEM tomographic software (D. Mastronarde, University of
Colorado at Boulder)
Miscellaneous ancillary equipment: e.g. a Haskris water circulator,
Zem water circulator; film desiccator; etc.
Auxilliary Equipment: (in the Bonner facility or in the nearby Natural
Sciences Building)
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Emitech K950X carbon evaporator equipped with a K150 film thickness
monitor and a K350 glow discharge device
Two manual, freeze-plunge devices for preparing vitrified samples
Folded optical diffractometer: used both for assessing quality of
micrographs and as a teaching tool to help new staff learn principles of
diffraction theory
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Nikon Super Coolscan 8000ED film scanner: for digitizing micrographs
at 4000 dpi pixel resolution.
Epson Perfection 4870 Photo flatbed scanner, 4800 X 9600dpi, 48bit
color depth
Two Dell Latitude D610 computers each with 1 GB memory, 80 GB
hard drive, 14 in flat panel displays, Linux Centos OS.
StorageWare SA214 2U Dual Xeon SATA server with one 80 GB SATA
hard drive and six 250 GB SATA hard drives
Hewlett Packard LaserJet 4350n printer, 1200 dpi
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