Vitrobot Mark IV System

Vitrobot Mark IV System offers reproducible vitrification of biological samples

The Thermo Scientific Vitrobot Mark IV System offers semi-automated vitrification to provide fast, easy, and reproducible sample preparation for cryo-EM. It performs the cryo-fixation process at constant physical and mechanical conditions like temperature, relative humidity, blotting conditions, and freezing velocity. This ensures high-quality cryo-fixation results and a high sample preparation throughput prior to cryo-TEM observation.

 

The Vitrobot System offers great value to the demanding scientific areas of cell biology and molecular imaging. It is equally suitable for food, industrial, pharmaceutical, and nanotechnological applications - where the true colloidal structure of your analyte is critical.

0:00

Vitrobot Mark IV System - sample vitrification for the Cryo-TEM workflow

Vitrobot Mark IV System automates the vitrification process to provide fast, easy, reproducible sample preparation - the first step in obtaining high quality images and repeatable experimental results.

Vitrobot Mark IV System features

The vitrification process

Highly automated vitrification process with enclosed process chamber:

  • Automated shutter control allows smooth, instant injection of the sample grid into the coolant (liquid ethane or propane). A lift for the container brings the coolant close to the shutter to ensure optimal vitrification.
  • Synchronous lowering of the coolant container and the grid holder keeps the grid submerged in the coolant and minimizes the risk of contamination prior to transfer ring the sample into a storage box or cryo holder.  
  • Coolant container with integrated anti-contamination ring.  
  • Grid transfer from the coolant towards a grid box in the liquid nitrogen environment is semi-automated. 
Cryo EM sample prepared using the Thermo Scientific Vitrobot Mark IV System

Sample application

Vitrobot Mark IV System provides precise and flexible control of all critical parameters in the plunge-freezing process:

  • Small sample volumes can be applied manually with a pipette through a small side port on the left and the right side of the climate chamber.
  • Both application time and wait time (between application and blotting) are software controlled and can be set in the user interface.
  • Precisely timed control of multiple sample applications, blotting actions, and vitrification enables time resolved analysis of interactions among separately applied components. 
Cryo EM sample application to a cryo EM grid with the Vitrobot Mark IV System

Vitrobot Mark IV System interface and instrument control

Easy and flexible user interface:

  • Linux operating system  
  • Touch screen control and set-up  
  • Mouse and foot pedal controls also available 
Vitrobot Mark IV screen interface

Vitrobot blotting device

  • Excess fluid is removed from the grid by (repeated) blotting with filter paper on rotating foam pads.
  • Number of blotting actions (max. 16 times for one grid) and duration of blotting are software controlled and can be set in the user interface.  
  • Longitudinal grid positioning (‘blot offset’) and wait time between blotting and vitrification (‘drain time’) are user definable. 
Thermo Scientific Vitrobot Mark IV blotting device

Vitrobot Mark IV System applications

The Vitrobot Mark IV System is compatible with multiple sample types, such as proteins, bacteria, and cells to fulfill your multiple structural biology applications requirements. 

Single particle analysis

Single particle cryo-EM analysis enables structural characterization at near-atomic resolutions from purified proteins or protein complexes.

Single particle analysis with cryo electron microscopy

Cryo electron tomography

Cryo-electron tomography is a technique that allows the study of the 3D structure of cells and tissues at near-native conditions.

Cryo electron tomography reconstruction of the Golgi apparatus

Integrative structural biology

Integrative structural biology combines mass spectrometry and cryo-EM for the determination of large dynamic complex structure.

Illustration of combined cryo-EM and mass spectrometry

How-to videos for operating the Vitrobot Mark IV System

Learn how to use the Vitrobot Mark IV System from preparing the system to vitrifying a sample with these step-by-step videos. 

Plunge-Freezing Tools
Review of the tools needed for plunge-freezing samples with the Vitrobot System.
Current Time 0:00
Duration 7:30
Loaded: 0%
Stream Type LIVE
Remaining Time 7:30
 
1x
    • Chapters
    • descriptions off, selected
    • subtitles off, selected
    • en (Main), selected
      1. Now Playing
        Up NextPlunge-Freezing Tools

        Review of the tools needed for plunge-freezing samples with the Vitrobot System.

      2. Now Playing
        Up NextVitrobot System connections

        Walk through of the Vitrobot System connections and cables.

      3. Now Playing
        Up NextMounting the Vitrobot humidifier

        Demonstration of how to properly mount the Vitrobot System's humidifier.

      4. Now Playing
        Up NextFilling the Vitrobot humidifier

        Step by step demonstration of how to properly fill the Vitrobot System humidifier.

    Tips and tricks for sample preparation on the Vitrobot System

    0:00

    Ask the Experts: Optimizing Samples for Successful Grid Preparation

    Learn common biochemistry and vitrification problems entotalsered during cryo-EM sample preparation and how to overcome them. Find out how to optimize and screen your samples prior to vitrification to help increase successful grid preparation.
    0:00

    Ask the Experts: Simplifying Cryo-EM with the Latest Innovative Tools

    In this webinar, we discuss the tools and technologies available as well as tips and tricks to help you get optimal results from highly difficult proteins for cryo-EM analysis.
    0:00

    Ask the Experts: Sample preparation and optimization for cryo-EM using the Vitrobot

    This webinar discusses tips and tricks for improving frozen hydrated cryo-EM sample preparation utilizing the Vitrobot. Frozen hydrated sample prep and optimization is often times the rate-limiting step in single-particle analysis.

    Vitrobot Mark IV System technical specifications

    Weight 31 kg
    Dimensions L/W/H: 413/260/890 mm

    Power supply
    • Voltage 110 – 230 V  
    • 50 – 60 Hz  
    • Fuse 4 AT (110 V USA)  
    • Fuse 2 AT (230 V Europe)  
    • Power cable 90 – 250 V 
    Operating parameters
    • Working temperature 4–60° C (at an ambient temperature range between 18–25° C)  
    • Peltier controlled heating/cooling  
    • Maintain relative humidity at 100%  
    • Ultrasonic controlled humidification

    Featured citations

    Find relevant citations from peer-reviewed journals where specific research results were generated using the Vitrobot Mark IV System.

     

    Cryo-EM grid optimization for membrane proteins

    Kampjut D, Steiner J & Sazanov LA. Cryo-EM grid optimisation for membrane proteins. iScience : 102139 (2021).

    https://doi.org/10.1016/j.isci.2021.102139

     

    Structure deformation and curvature sensing of PIEZO1 in lipid membranes

    Yang, X., Lin, C., Chen, X. et al. Structure deformation and curvature sensing of PIEZO1 in lipid membranes. Nature 604, 377–383 (2022).

    https://doi.org/10.1038/s41586-022-04574-8 

     

    Integrative structure of a 10-megadalton eukaryotic pyruvate dehydrogenase complex from native cell extracts.

    Kyrilis FL, Semchonok DA, Skalidis I, et al. Integrative structure of a 10-megadalton eukaryotic pyruvate dehydrogenase complex from native cell extracts. Cell Reports 34 (2021).

    https://doi.org/10.1016/j.celrep.2021.108727 

     

    Dynamics of GLP-1R peptide agonist engagement are correlated with kinetics of G protein activation.

    Deganutti, G., Liang, YL., Zhang, X. et al. Dynamics of GLP-1R peptide agonist engagement are correlated with kinetics of G protein activation. Nat Commun 13, 92 (2022).

    https://doi.org/10.1038/s41467-021-27760-0 

     

    Structural basis of actin filament assembly and aging

    Oosterheert, W., Klink, B.U., Belyy, A. et al. Structural basis of actin filament assembly and aging. Nature 611, 374–379 (2022).

    https://doi.org/10.1038/s41586-022-05241-8

    Cryo-EM sample preparation and sample analysis additional resources

    Cryo-EM Sample Preparation

    Successful protein preparation lies at the base of any successful structural biology technique. For cryo-electron microscopy, a wide range of solutions is required to achieve the highest sample quality before freezing of the specimen on the EM grid. Find solutions from protein expression, preparation and vitrification. 

    Successful protein preparation

    Cryo-EM Resource Center

    This resource hub contains rich and reliable technical content designed for new and experienced researchers who are exploring the capabilities of cryo-EM. 

    Successful protein preparation

    For Research Use Only. Not for use in diagnostic procedures.