Titan Krios - Changing Energy

Cryo-EM Setup for Changing Energy

Quick Reference for Changing Energy on the Titan Krios ❄️

Titan Krios - Changing Energy

Cryo-EM Setup for Changing Energy

Quick Reference for Changing Energy on the Titan Krios ❄️

Overview

There are three main places that energy settings must be imported:

  1. Microscope Energy
  2. Microscope Alignments
  3. FEG Registry
  4. Gatan Settings


Step 1 - Microscope Energy

  1. Close the column valves.
  2. Close any instance of EPU that is running.
  3. Set the energy. Setup Tab >> High Tension >> 200 kV. If you expand the High Tension tab, you will see the high tension move to 200 kV and stabilize.

Step 2 - Load Alignments

  1. Open the Alignments tab.
  2. Select the appropriate alignments file (e.g. the most recent 200 kV settings).
  3. Available settings will propagate in the "available" window. Select all of them by clicking on one, then hitting Ctrl+A. Click the < arrow to move the selected parameters to those being used.
  4. Click Apply.
  5. Wait overnight for the microscope to stabilize before collecting data. Some preliminary work can be done once the energy has stabilized.
  6. Perform a quick beam center. Lift the screen. Direct Alignments >> Beam Shift, adjust with Multifunction X and Y on the control pad, then click Done on the Direct Aligments tab.

Step 3 - FEG Registry

  1. Set the FEG Controls. FEG Control >> Gun Lens. Set the gun lens (5 for 200 kV; 4 for 300 kV). Confirm that the settings have been applied in the scope settings window.
  2. Load the FEG Registry. FEG Register. Select the appropriate registry (e.g. the most recent 200 kV settings). Click Set.

Step 4 - Setup Gatan Camera

  1. Activate the Filter Control Menu.
    1. Help >> User Mode >> Power User.
    2. Window >> Filter Control
  2. Save the current settings. Look for the 💾 icon near the bottom of the Gatan Filter Control window.
  3. Change the energy as appropriate.
  4. Load the settings (📂 icon near the bottom of the Gatan Filter Control Window).
  5. Center the ZLP.
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Jonathan Ipsaro
Research Specialist

Biochemist with a penchant for integrated structural, biophysical, and computational methods.

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