Structural CryoEM

Structural electron microscopy facility

The Department of Pharmacology, Physiology and Biophysics in the Center for Advanced Biomedical Research (CABR) houses a pair of cryogenic capable, electron microscopes for use in visualizing biological nano-structures in negative stain or frozen vitreous buffer. The Structural EM suite consists of 4 rooms in total (~900 square feet) with sample preparation room and a grid freezing room; the latter is maintained as a contiguous dry space with the microscope rooms (~8-15% RH), with a Harris Environmental Systems humidity control system. For grid freezing, Vitrobot Mark 3 and Mark 4 plunge freezing robots are available to all Departmental users along with a Nanosoft grid mounting/clipping station with stereo dissecting microscope and cryotools to prepare grids for insertion and visualization on automated ThermoFisher electron cryo-microscopes equipped with autoloaders. An outer support room houses a Denton carbon evaporator, a glow discharge unit, a Gatan dual cold holder pumping station and cryo-storage dewars.

Conventional cryo-TEM “bubble” microscopes –

CM12: A cryo-CM12 with dedicated Gatan 60º cold stage is available for Departmental users and for cryo-EM training. The CM12 has computer-controlled stage position and relocation software/hardware with an upgraded external computer to run the low dose kit. The microscope is equipped with a LaB6 filament and a 2k x 2k TemCam-F216 camera (TVIPS) for cryo-imaging.

Tecnai F20: The Structural EM Facility contains a Tecnai F20, a Field Emission Gun electron microscope for cryogenic imaging with a 4k X 4k TemCam 416 CMOS camera (TVIPs) with separate imaging PC and monitor running EM-MENU4, EM-tools, and Serial-EM. The CMOS camera and software are used for automated data collection for single particle analysis, tomography and to optimize samples for data collection on automated cryo-electron microscopes at National cryo-EM beamlines or in the new Boston University cryo-EM core facility, located on the Medical School Campus. There are 2 dedicated Gatan cold holders (DH676), 60 and 70 degree tilt, and an Oxford cryo-holder for the TF20. The microscope PC has been rebuilt to provide a stable system for imaging and data collection.

Cryo-capable electron microscopes in the Structural EM facility including the CM12 (top left) and Tecnai F20 (right and lower panels).
Vitrobot Mark 3 (top) and Mark 4 (bottom right) grid plunging robots with accessories, including a Nanosoft grid mounting station(bottom left).
General equipment including: Gatan pumping station and storage dewars (top), Denton carbon evaporator and glow discharge instrument (bottom left). A 6 node Linux cluster with 192 CPU, 24 GPU and a 50 Tb RAID is located in an adjacent server room and used for single particle and tomographic data processing (Akey research group, bottom right).

Department EM user pages:

Christopher Akey
Esther Bullitt
William Lehman
Clint Makino
Olga Gursky

Cryogenic EM Core Facility