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After screening your grids start an EPU session.
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Make sure to tick option 'Tilted Acquisition'.
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Similarly to a regular EPU session - select the grid squares on which you would like to collect your data.
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Specify the stage tilt angle, typically 30 or 40 deg.
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Click 'Auto Eucentric'.
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Measure hole size - please note that due to the stage tilt the hole appear oval rather than circular. The two yellow patterns adopt the oval shape accordingly.
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Press 'Remove Holes Close to Grid Bar', prepare all squares and clean them with the ice filter and/or selection brush.
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Import your data collection template.
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Rearrange the green data acquisition areas to ensure data collection over the ice rather than carbon.
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Click 'Show/Hide Tilt Axis'. Tilt axis will be displayed as blue line. On the Glacios G2 it will be ~vertical (please see the corresponding screenshot), on the Krios G3i it will be ~horizontal.
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Position 'Autofocus Area' on the tilt axis.
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In the microscope user interface (left screen, bottom-right corner) the alpha tilt (A) will show the current stage tilt (30 or 40 deg, depending on your settings).
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Open the 'Stage' control panel by clicking the menu in the bottom-right corner.
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Enter value '0' next to 'Set Alpha'.
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Click 'Set Alpha' - the button will become yellow and the alpha tilt (A) will change to '0'.
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Unclick the 'Set Alpha' button. It should turn gray again, meaning the option is deactivated.
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Proceed to direct alignments, as usual.
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Once the EPU run has started the stage will be automatically tilted to the preset angle.
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