CryoEM

Services

Located in the Morrison Center, the CryoEM Core Facility has a stage-of-the-art Glacios 200kV Electron microscope available for our community for structure determination of protein, nucleic acids, and other molecules of biological interest, including small molecules in a crystalline state.

Our Services include:

• Single-Particle CryoEM (SP-CryoEM).
• Cryo-Electron Tomography (CryoET) of Thin specimens.
• Micro-Crystal Electron Diffraction (MicroED). 
• Sample Preparation.
• Comprehensive Data Collection and Analysis of CryoEM samples.
• Sample Storage and Shipping.
• Data Processing. 

Hours: Monday-Friday 9:00 am-5:00 pm for regular users, and 24h access for advanced users.

The CryoEM core facility is supported by Federal funding by NIH COBREs (NCIBC) and usage/service fees. 

Service Fees

1. New Users are Required to Train for at least 8 h before using the instrument unsupervised.
2. The Minimum Reservation time for the Glacios is 4h starting at 9 am or 1 pm.
3. The Vitrobot and Leica vitirification robots can be reserved in blocks of two hours.  Training is required for unsupervised use.
4. The Glacios CryoTEM have three types of bookings Glacios Screening, Glacios Overnight, and Glacios per Day.
5. Glacios Overnight can only be used by trained users.
6. A glow discharger fee will be added to all the vitrification robots reservations.

Other consumables will be charged at cost. Please reach out to the Facility Director if you require a quote for our services.

Instruments

200kV Glacios CryoTEM

The Thermo Scientific Glacios Cryo-Transmission Electron Microscope (cryo-TEM) allows you to easily collect near atomic data from a broad range of biological targets. Compared to the previous generation, the Glacios Cryo-TEM delivers higher throughput and makes cryo-EM more accessible and efficient, thanks to the AFIS and FFI systems. The Glacios features a Falcon 4i Direct Electron Detector, a Selectrics Energy Filter, and the Thermo Scientific EPU Software. In addition, the instrument is capable of loading up to 12 grids reducing sample contamination and speeding up the screening process. Furthermore, Our Glacios has the MicroED package, which includes a CetaD camera, optimized apertures, a smaller beam stop, and EPU-D. The Glacios Cryo-TEM is ideally suited for Single-particle Analisis, Cryo-Electron Tomography (Cryo-ET), and microelectron diffraction (MicroED). To reserve our Glacios, please click here, or contact the facility Director.

200 kV Glacios CryoTEM

TFS Vitrobot Mark IV

The Vitrobot is a fully computer-controlled double-sided Bloating Robot for the vitrification of aqueous samples. Minimal training is required for operating the instrument in a laboratory environment. The process of plunging, blotting, and vitrification is fully automated upon placing a vial in the chamber and setting up the liquid coolant container. Operational parameters tend to be reproducible, enabling a high throughput of vitrified samples with a straightforward control of the vitrification process. The Vitrobot allows you to control the temperature, humidity, number of blotting, and several critical time settings. The instrument will consistently produce excellent specimens for Cryo-electron microscopy, including single-particle, CryoET, and MicroED. To reserve our Vitrobot, please click here, or contact the facility Director.

Vitrobot Mark IV

Leica EM GP2

The Leica EM GP2 is a fully computer-controlled single-sided Bloating Vitirifcation Robot. Bloating on the EM GP2 can be performed using sensor bloating or manual blotting using the onboard eyepiece. This instrument allows the user to perform multiple bloating operations, including back and front bloat. The possibility of back bloating makes the EM GP2 a perfect tool for the vitrification of cells and microcrystals. In contrast with the Vitrobot, the EM GP2 is capable of controlling the temperature of the liquid ethane and providing a continuum flow of liquid nitrogen in the sample vitrification area, considerably reducing ice contamination. The instrument is highly reproducible and excellent for the three main CryoEM workflows, including single-particle CryoEM, CryoET, and MicroED. To reserve our Leica EM GP2, please click here, or contact the facility Director.

Leica EM GP2

PELCO easiGlow

The PELCO easiGlow™ Glow Discharge Cleaning System is a compact, quick, easy-to-use standalone system. It is designed for cleaning and hydrophilizing TEM carbon support films or grids, which tend to be hydrophobic. A glow discharge treatment with air will make a carbon film surface negatively charged (hydrophilic), which allows aqueous solutions to spread evenly. Glow discharge treatment of TEM grids removes adsorbed hydrocarbons, cleaning them while making them hydrophilic. The PELCO easiGlow™ supports hydrophilic and hydrophobic treatment with either negative or positive charge and includes two separately controlled gas inlets. The vacuum level in the instrument is precision controlled electronically by a proportional valve, a novel approach that eliminates manual settings with a needle valve. A gentle venting procedure ensures that the TEM grids are not disturbed when the system is vented.

PELCO easiGlow

Reservations 200kV Glacios CryoTEM

Check the calendar below and then submit a request  to use an instrument a minimum reservation time of 4h is required.

In order to submit a glacios overnight or Glacios per Day request, your grids needs to be screened prior to your data collection.

If you are having issues viewing this calendar, simply try to refresh the page.  If the issue persists, you may need to try another browser.

Please note:  If users arrive late, the billing time will start at the beginning of the scheduled appointment. Users are required to check in and out of the microscope by completing the following Google form or scanning the QR code located in the control room.

Reservations Vitrobot and Leica EM GP2

Check the calendar below and then submit a request to use an instrument a minimum reservation time of 2h is required.

If you are having issues viewing this calendar, simply try to refresh the page.  If the issue persists, you may need to try another browser.

Please note: If users arrive late, the billing time will start at the beginning of the scheduled appointment. Users are required to check in and out of the microscope completing the following google form or scanning the QR code located in the instrument proximity.

If you are planing to clip your grids please reserve the clipping station here.

Clipping station booking

Check the calendar below and then submit a request to use an instrument a minimum reservation time of 1h is required.

If you are having issues viewing this calendar, simply try to refresh the page.  If the issue persists, you may need to try another browser.

Please note:  Users are required to check in and out of the instrument by completing the following google form or scanning the QR code located near the instrument.

Policies

• Clipping Guidelines
• Vitrobot Guidelines
• Leica EM GP2 Guidelines
• Glacios Guidelines

Personnel

Research/Service Group

Dr. Eduardo E. Romero, Director
Morrison 140
Phone: 402-472-3360
Email Eduardo

Publications

Advisory Committee

Facility Advisory Group
The mission of the Facility Advisory Group is to provide guidance and feedback to the facility regarding the most effective use of its resources to meet research demands.

Contact the Director

Director: Dr. Eduardo E. Romero
Office 140, Morrison Center, East Campus

Telephone Number:
402-472-3360 (Office)
Email: eduardo.romero@unl.edu