Apreo FE-SEM Basic Training xT Microscope Control & Operation Central Analytical Facility The University of Alabama

Microscope Server and User Interface (UI) While the xT Server software should be open at all times, to start it double click the icon Click the “Start” button Mouse over the green bar and double click to expand the server menu Do not click “Stop” as this will stop the server Login info: Name: User PW: User Name: Supervisor PW: Supervisor

xT Microscope Control Tool Bar Image Windows Pages and Menus Data Bar Navigation Cam CCD Objective Lens ETD/SE Detector

Menu Bar – File & Detectors Open files Save & Save As Save all Saves an image from each quadrant Record Movie Print Import & Export parameters List of detectors, used to change image windows ETD (secondary e-) T1 (BSE) T2 & T3 (secondary e-) CCD Live for stage movement Nav Cam Double click image to move to desired area

Menu Bar - Scan Pause (F6) Snapshot (F4) Snapshot (F2) Videoscope (F3) Slow scan with prompt to name and save image Snapshot (F2) Slow scan that automatically saves to selected folder Videoscope (F3) Histogram displaying signal, used for brightness/contrast adjustment Reduce Area (F7) Smaller display window for focus and stigmation Beam raster options Scan speed controls Image Filters Live image Average – Averages # of live scans to produce picture Integrate – Averages # of live scans to produce picture, pauses scanning Scan Rotation Preferences

Menu Bar – Stage Align Feature Compucentric Rotation Define User Units Aligns a feature to either the horizontal or vertical axis through stage rotation or scan rotation Compucentric Rotation X & Y axes are moved during rotation to keep ROI centered in image window Define User Units Beam Shift Reset Centers beam shift, also available on beam page Home Stage Drives stage to home position with or without rotation Link/Unlink Z to FWD Nav-Cam options Can drive to the Nav-Cam position for an updated photo as needed, or restore the previous photo

Menu Bar – Tools & View Center Cross Auto Contrast/Brightness (F9) Auto Focus (F11) Auto Stigmator (Ctrl + F11) Center Cross

Menu Bar - Help DocumentationPDF version of the instrument manual. Can also be accessed by clicking on and “?” icons displayed in the software Safety Manual Keyboard Shoftcuts A list of F key functions and other shortcuts for operation User Guidance Workflows for specific applications, Useful for suggestions on beam parameters, working distance, etc.

Tool Bar 1) Undo/Redo buttons 2) Sample exchange window 3) Source selection (e-, Nav-Cam, CCD) 4) Use case selection (Standard, Optiplan, Immersion) 5) Dropdowns for magnification, accelerating voltage, and spot size/current 6) Lens adjustment 7) Reduced area (F7) 8) Link Z to FWD Makes Z distance a function of the working distance from objective lens to sample surface 9) Auto contrast/brightness, focus, and stigmation

Tool Bar 1) Auto lens adjust 2) Videoscope (F3) 3) Dropdowns for dwell time and image resolution 4) Snapshot (F4) 5) Photo (F2) 6) Pause (F6) 7) Image Filter (Live, average, integrate) 8) Measurement and annotation tools

Beam Page Pump/Vent controls Column controls Magnification drop down “Beam On” will open/close the column valve, exposing the sample to the e- beam Dropdowns for Spot Size/Current or High Voltage Magnification drop down Compound Lens Filter Used in conjunction with Immersion mode. Involves applying a magnetic field between objective and sample surface at very small working distances to form the smallest probe possible Beam Options An alternative adjustment for stigmation and beam shift. Right click to zero these parameters if needed Scan Rotation Detector contrast and brightness slider bars

Stage Page Coordinate system Positions table Stage Nav-Cam view Actual – Current coordinates Target – Coordinates to move to Relative – Coordinates relative to actual Positions table Save, update, and remove stored positions Stage Nav-Cam view Depiction of stage relative to door and ETD Stage Z slider bar Fine adjustment of the stage in the Z axis Scan rotation For image rotation when stage rotation is not an option

Venting Chamber and Loading Samples After ensuring the column valve is closed, select vent from either the sample exchange window or the beam page. The venting process will take ~ 80 seconds During venting the stage will be automatically lowered Once vented, grab the chamber door handle and open the stage to load/unload samples Note that only positions on the stage marked with “screw” need a set screw. Positions in the center of the stage do not Be mindful of sample height so as to not damage the objective lens when closing the chamber door From either menu select the “Pump” option to evacuate the chamber If the “Take Nav-Cam Photo” box is checked, the stage will drive to the Nav-Cam for an updated picture

Coarse Focus, Linking Z to FWD Once target pressure is reached (<7.5 x 10-6 torr), open the column valve by selecting “Beam On” in the Beam page Using the coarse and fine MUI focus and stigmator knobs, focus on the surface of a sample up to ~10k X magnification Once focused, click the “Link Z to FWD” button The Z coordinate displayed in the actual coordinate set on the Stage page is now a function of distance between the objective lens and the sample surface Focus and stigmator knobs on MUI

Moving the Stage and Fine Focus There exist three ways to move the stage: 1) Typing in a target coordinate after linking Z to FWD 2) Using the Z slider bar 3) Holding down the mouse wheel will change the mouse icon to a yellow bar. Moving the mouse forward or backward will raise/lower the stage. Also works in imaging window for X & Y movement Raise the stage to the 10 mm position (yellow line) Repeat focus/stigmation at a higher magnification As a general rule, it is a good idea to focus/stigmate at 1.5-2 times the magnification you wish to capture an image at Link Z to FWD one additional time This process should repeated when moving between samples, especially those with drastically different heights

Saving images After repeating the focus/stigmation process to produce an image, there are multiple ways to scan and save said image Snapshot (F4) and Photo (F2) will increase the dwell time of the beam before saving the picture. This can help remove some of the graininess in the image Average or integrate a desired number of frames before using the “Save” or “Save As” functions. This can be useful at higher magnifications where vibration is a factor, and a slow scan may not remove it “Save All” will save images from any of the four quadrants, with check boxes available for each as well as picture formats available in dropdowns

Shutdown When imaging is complete, the instrument will need to be put in a shutdown stage for the next user Begin by closing the column valve (“Beam On” on the Beam Page. Icon is grey when valve is closed, yellow when open) Lower the stage a sufficient distance to prevent collision with the objective lens upon opening the chamber. 35-40 mm is a good target. Turn off and features such as Beam Deceleration or the Compound Lens Filter Return the lower left and lower right quadrants to the Nav-Cam and CCD respectively, using the Detectors menu Either through the Beam Page or Sample Exchange Window, select “Vent” to begin venting the chamber. This will take ~80 seconds. Once vented, use the handle on the front of the chamber to open the chamber and remove samples If the next user is present the chamber can be left vented for them to load samples. If the next user is not present press “Pump” to begin evacuating the chamber Data can be retrieved from the support PC located in the Data Processing room