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 came 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 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. The operating system was upgraded from Windows XP to Windows 7 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) BioQuantum
• Post-GIF  Gatan K3 camera
• Volta Phase Plate
• Pre-GIF Falcon II detector
• Pre-GIF Gatan Orius CCD (soon to be replaced with an FEI Ceta camera)

Main applications

• Single particle cryoEM
• Electron cryotomography

Janelia Krios 2

This microscope was set up in 2013 and has been upgraded to stay at the cutting edge. 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 March 2019 the TVIPS camera was replaced with a GIF BioQuantum and K3 camera. The operating system was upgraded from Windows XP to Windows 7 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
• Post-GIF  Gatan K3 camera
• Volta Phase Plate
• Pre-GIF Gatan K2 camera

Main applications

• Single particle cryoEM
• Electron cryotomography

Janelia Krios 3

Main configuration

• High-brightness field emission gun (X-FEG)
• Energy filter: Gatan Image Filter (GIF) BioQuantum
• Post-GIF  Gatan K3 camera
• Volta Phase Plate
• Pre-GIF Falcon 4 camera
• Pre-GIF Ceta camera
• Scanning Transmission Electron Microscopy (STEM) capability

Main applications

• Single particle cryoEM
• Electron cryotomography
• STEM tomography

FEI Aquilos Cryo FIB

The system will be upgraded to Aquilos 2 in late 2020 for improved ease of use, productivity, automation, and capability to lift a slab from a frozen bulk sample for FIB milling.

Main application

• Sample preparation platform for electron tomography
• Use focused ion beam (FIB) to mill down interested region of frozen cells into thin cell lamella (150-300 nm thick)
• Scanning electron microscopy (SEM) capability

Leica Cryo CLEM light microscope

Autogrids can be used directly on the system that can be later transferred to the Aquilos cryoFIB. The system will be upgraded into a Leica Thunder cryo light microscope in 2020.

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.

TTP 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.