Equipment

Janelia Krios 1

This microscope was set up in 2011. It has gone through multiple rounds of hardware and software upgrades to keep up with new developments in the cryoEM field. This Krios was one of the few in the world that is equipped with a Cs corrector to reduce spherical aberration of the objective lens from 2.7 mm to ~0.01mm. It came with a film camera, which is still on the scope. At the time of purchase direct detectors were still in early days of development and films were still in use. The other original cameras that came with the microscope were a pre-GIF Falcon I and a post-GIF Gatan US4000 CCD. Later the US4000 CCD was replaced with a Gatan K2 Summit camera and the Falcon I was replaced with a Falcon II camera. In September 2019, the post-GIF K2 camera was replaced with a Gatan K3 camera. In December 2022, the Gatan Quantum energy filter was replaced with a Gatan BioContinuum HD filter. The operating system was upgraded to Windows 10 and the main microscope operating software and data collection software have been frequently upgraded.

Main configuration

• High-brightness field emission gun (X-FEG)
• Spherical aberration corrector (Cs corrector)
• Energy filter: Gatan Image Filter (GIF) BioContinuum HD
• Post-GIF  Gatan K3 camera
• Volta Phase Plate

Main applications

• Single particle cryoEM
• Cryo electron tomography

Janelia Krios 2

This microscope was set up in 2013 and has been upgraded frequently. It came with a standard field emission gun (s-FEG) which was later upgraded to an X-FEG. This Krios was originally configured for single particle data collection and electron diffraction. There was no GIF and the main camera set up was a Gatan K2 Summit detector directly under the projection chamber followed by a TVIPS F416 camera. In  2019 the TVIPS camera was replaced with a GIF BioQuantum and K3 camera. The operating system was upgraded to Windows 10 and the main microscope operating software and data collection software have been frequently upgraded.

Main configuration

• High-brightness field emission gun (X-FEG)
• Energy filter: Gatan Image Filter (GIF) BioQuantum
• Pre-filter Gatan Oneview camera
• Post-GIF  Gatan K3 camera
• Volta Phase Plate

Main applications

• Single particle cryoEM
• Cryo electron tomography

Janelia Krios 3

• Cold field emission gun (C-FEG)
• Energy filter: Thermo Scientific Selectris X
• Pre-filter Ceta camera
• Post-energy filter Thermo Scientific Falcon 4i camera 
• Volta Phase Plate

Main applications

• Single particle cryoEM
• Cryo electron tomography

FEI Aquilos 2 Cryo FIBSEM

Main application

• Sample preparation platform for cryo electron tomography
• Use focused ion beam (FIB) to mill region of interest on frozen hydrate cells into lamella (100-200 nm thick)
• cryo lift-out capability to work on frozen hydrate tissue sample

FEI Hydra cryo plasma FIBSEM (cryo pFIBSEM)

A Hydra cryo pFIBSEM was ordered and is estimated to be delivered in the fall of 2023. 

Leica Cryo CLEM light microscope

Autogrids can be used directly on the system that can be later transferred to the Aquilos2 cryoFIB. 

Main application

• Identify regions of interest in cells frozen on a cryo-EM grid using wide field and/or fluorescence light microscopy that may be used later to aid cryoFIB milling.

STP Labtech Chameleon System

This is a new generation cryo-EM grids preparation equipment. It dispenses a small volume (a few nanoliters) of sample onto a self-wicking grid while the grid is being plunged. For certain samples it may help reduce negative effects at the air-water interface.

Plunge freezers and screening TEMs

The facility also hosts multiple plunge freezers and other small equipment for making cryoEM grids. A Tecnai Spirit Biotwin TEM and a Tecnai F20 TEM  can be used to image negative stain grids or to screen cryo grids.