Cryo-electron microscopy and cryo-electron tomography core facility

The Rutgers New Jersey Cryo-Electron Microscopy and Tomography Core Facility images and creates high-resolution three-dimensional maps of biomolecules preserved in native or near-native conditions by rapid vitrification. Our versatile setup can be used for high-resolution experiments as well as analysis of more heterogeneous or pleomorphic biospecimens. We provide training to students, staff, and faculty to disseminate knowledge in this field.

The facility is available on a fee-for-use basis, with preference in scheduling and pricing to members of the Rutgers community. Potential users are welcomed to contact the director to discuss how cryo-electron microscopy or tomography can be applied to their projects. Before bringing a specimen to the facility, users should provide supporting data that their specimen is of a suitable concentration (generally, 0.2-1 mg/mL) and of sufficient purity for cryo-electron microscopy.

Equipment

Our 200 kilovolt Talos Arctica cryoelectron microscope represents the state-of-the-art in optics and stage technologies. It is equipped with a BioQuantum Gatan Image Filter to remove inelastically scattered electrons, a Volta phase plate for contrast enhancement, and a Summit K2 direct electron detector for optimal detector performance at high resolution. This configuration is suitable for both single-particle and tomographic studies. The instrument is piloted remotely from an adjoining control room outfitted with automated collection software.

Vitrification is achieved with a Leica EM-GP plunge freezer, capable of front- or back-blotting. Plunge freezers generate an aqueous layer of reproducible thickness across the EM grid and rapidly chill this layer to an amorphous, solid state. The EM-GP is ideal for cell studies and also creates uniform-thickness ice for single-particle work. We stock standard grid styles and have a Pelco EasiGlow available for hydrophilization.

Correlative light microscopy/electron microscopy (CLEM) experiments are conducted on a Leica DM6 FS/EM Cryo CLEM system. After vitrifying a sample on an electron microscopy grid, the same grid can be imaged in fluorescence (or brightfield) optical microscopy and in the cryoelectron microscope. Fiducial markers seen in both optical and electron microscopy are used to correlate the images. In one embodiment, a quick scan of the grid by optical microscopy is used to locate a rare event, and the user tracks the electron microscope stage to the correct site to record a tomogram at that location

The facility maintains some resources for data processing in-house on a buy-in basis. We can also assist in setting up processing software or workflows on computing clusters.

Useful Links

Cryoelectron microscopy is complementary to classical electron imaging techniques such classical transmission electron microscopy. The Core Imaging Lab, a unit of the Robert Wood Johnson Medical School Department of Pathology, provides services such as TEM of stained or sectioned samples to the Rutgers community.

The research interests of several members of the Institute for Quantitative Biomedicine include structural biology generally or cryo-electron microscopy techniques. In particular, Professors Wei Dai and Arek Kulczyk are leaders in cryo-electron tomography and cryo-electron microscopy at Rutgers. For example, the Kulczyk lab is developing new techniques in single-molecule CLEM as well as applying high-resolution single-particle analysis to biological targets. The Dai lab integrates cryo-electron tomography with CLEM and other tools to study nanomachines and disease-associated structures in their native cellular context, and has pioneered application of phase plates in biological electron microscopy.

Please see the IQB Bootcamp page to find information about the upcoming weeklong winter boot camp on cryo-electron tomography.

Contact information

The facility is located on the ground floor of the Proteomics building, which we share with the high-field NMR and biological mass spectrometry facilities. The facility director, Prof. Jason Kaelber, can be contacted by email, and at 848-445-5302 on occasions when he is not in the experimental suite.