Leica S430 Scanning Electron Microscope Operations Checklist

From Pumping Station: One Wiki
Jump to: navigation, search


The SEM Operation checklist in Section 2 contains step by step instructions to operate the SEM. Note that these steps reference Procedures described in PROCEDURES. These are listed in alphabetical order. For troubleshooting, see TROUBLESHOOTING.



  • Verify power switch in Standby, vacuum pumps running.
  • Power switch to On.
  • Enter BIOS Password.
  • Run LEO UIF.
  • Select user directory.
  • View -> DataZone.
  • Check vacuum.
  • No vacuum / right pulldown says At Air:
  • Vacuum in e-4 torr range:
  • Vacuum in e-5 torr range or better: Proceed.
  • Set X to 50.00
  • Set Z to 31.0 (or less if a tall sample is in the chamber)
  • Set Tilt to 0


  • Press Stoplight icon.
  • Wait for image.
  • Adjust Brightness. (Contrast / Brightness icon)
  • Set EHT appropriate to sample.
  • Locate sample.
  • Focus. (see FOCUS procedure)
  • After 5 minutes since adjusting EHT have elapsed, follow FILAMENT CURRENT procedure.
  • If an aperture other than #2 is required, follow CHANGE APERTURE procedure. NOTE: #2 aperture is usually the best aperture, so changing apertures isn’t necessary.


  • Move sample as required to center item of interest.
  • Zoom (Magnifying glass icon, left mouse button) as required.
  • When zooming, stop to focus (see FOCUS procedure) frequently.
  • At very high magnification, move field of view with beam shift:
    • Beam -> Stigmation..
    • Click Beam Shift.
    • Drag scroll bars or position dot as needed.
    • Press HOME key to undo all beam shift.
    • Consider noise reduction (see NOISE REDUCTION procedure) if necessary prior to exporting images.
    • Export images as needed. (see EXPORT IMAGE procedure)


  • Change to aperture #2 and align aperture if you previously changed to a different aperture. Follow CHANGE APERTURE procedure.
  • Set X to 50.00
  • Set Z to 31.0 (or less if a tall sample is in the chamber)
  • Set Tilt to 0
  • Single click upper left box.
  • Click SEM Shutdown.
  • Shutdown: Save Operating Conditions? Click No.
  • Follow PUSH IMAGES procedure if you saved images.
  • Double click upper left box of Program Manager.
  • Click OK.
  • At DOS prompt, click Standby power button.



  • Zoom in to 1000x.
  • Center on noticeable object or feature.
  • Follow FOCUS procedure, including anti-hysteresis. (F2)
  • Reduced raster (Monitor icon with square) - size and drag to cover noticeable object or feature in question.
  • Scan rate to fastest. (Click Fast Forward icon 3 times)
  • Click Focus Wobble icon. (Icon with red half sine wave at bottom)
  • Adjust X and Y aperture controls, one at a time, to cancel movement.
  • Click Off button in focus wobble box.


  • Beam -> Apertures.
  • Select aperture number.
  • Click OK.
  • Turn Y aperture control to select proper aperture:
  • Turn knob IN for 1, OUT for 3
  • E.g. to change from aperture #2 to #1,turn clockwise
  • Adjust Y for maximum brightness.
  • Follow ALIGN APERTURE procedure.


  • File -> Export TIFF
  • Click Freeze if image not already frozen.
  • Click OK to export.
  • Image -> Unfreeze


  • Scan rate to fastest. (Click Fast Forward icon 3 times)
  • Click light bulb icon.
  • Left mouse button – adjust I(M) for 1st peak.
  • Lower Brightness (Contrast / Brightness icon) if too bright to differentiate peak, then continue adjusting filament current.
  • WARNING: Increasing filament current substantially past 1st peak shortens filament life.
  • NOTE for 2nd peak operation:
  • Possible IF AND ONLY IF you can identify a well-defined 1st and 2nd peak.
    • Increase I(M) current SLOWLY. Note that a falling off after 1st peak and sharp rise to 2nd peak should be identifiable when increasing filament current.
    • If these features are not visible within 0.1A to 0.2A beyond 1st peak, STOP. DECREASE FILAMENT CURRENT and operate at 1st peak only.
    • If well defined 2nd peak is visible, adjust for maximum brightness such that ANY decrease in current decreases image intensity. NOTE: Any operation above this point even more drastically shortens filament life and must be avoided.


  • Consider anti-hysteresis (F2) after major parameter changes. (zoom changes
  • Consider reduced raster. (Monitor icon with square, size and drag as appropriate)
  • Set appropriate scan rate. (Fast Forward / Rewind arrow icons)
  • Select Focus. (Magnifying glass icon)
  • Middle button: drag to set Working Distance and focus image.
  • Full screen raster. (Monitor icon without square)
  • Set appropriate scan rate.
  • NOTE: The image noticeably shifting during focus adjustment indicates the aperture is not in alignment. Follow #ALIGN APERTURE procedure.
  • NOTE: Auto-focus can be done via F key but it can sometimes make focus worse.


  • Scan rate to slowest. (Click Rewind icon 3 times)
  • Image -> Noise Reduction..
  • For Frame Mode, which scans at the same rate but averages multiple successive frames, making the image clearer:
    • Increase “Frames to average” slider.
    • Click Frame Avg. button.
  • For Line Mode, which scans each line repeatedly and averages each line:
    • Increase “Line Avg.” slider.
    • Click Line Avg. button.
  • After making adjustments, wait for scan to pass the bottom of the screen and, when visible at the top, click Freeze mode / At end button.
  • When blue square appears in the lower right corner of the screen, the image is frozen and can be exported. Follow EXPORT IMAGE procedure.
  • Restore both sliders to the left and click Frame Avg. button.
  • Close Noise Reduction window.
  • Image -> Unfreeze.


  • WARNING: Only do this when LEO UIF software is shut down!
  • Double click “Push [Your Name]” from list of icons in User Operations window.
  • Black screen and lots of scrolling text: Success.
    • Download images from ftp://semsrv.pumpingstationone.org to a laptop or other device. NOTE: This URL is only available on PS:One's internal network.
    • Double click “Delete [Your Name]” from list of icons in User Operations window. This deletes the images from the SEM hard disk.
    • NOTE: No automatic delete happens on the SEM hard disk. Users must delete images once they have been safely transferred off the SEM.
    • Images will be deleted automatically from the server in two weeks.
  • OR black screen and one line of text, and everything has paused: Failure. NOTE: This failure mode is fairly common after operating the SEM.
    • Hit Ctrl-Alt-Delete. At blue screen, hit Enter.
    • Shut down Windows as usual.
    • At the DOS prompt, Ctrl-Alt-Delete to reboot.
    • Enter BIOS Password.
    • Repeat this procedure WITHOUT LAUNCHING LEO UIF.


  • Press V (if vacuum window not open).
  • Click Pump.
  • OK to start beam once vacuum stabilizes at or below 6e-5 torr.
  • Close Vacuum window.
  • If vacuum does not reach this level in 20 minutes, report to mailing list.


  • Press V (if vacuum window not open).
  • Click Vent.



  • In case of erratic mouse movement:
    • Lift mouse, shake it, blow into track ball
    • Shift the cord around.
  • If mouse is stuck changing something (magnification, focus, brightness, contrast, etc.) when the button is no longer held down:
    • Press Escape.
    • Click anywhere else within the image.


  • If Off light is illuminated:
    • SEM likely lost power. Report loss of power to mailing list.
    • Press Standby.
    • Wait 30 seconds.
    • Press On.
    • Proceed as normal, but follow PUMP CHAMBER instructions.
  • If no lights illuminated:
    • If the vacuum pump is running:
      • Bulb likely burnt out. Report to mailing list.
      • Press On, proceed as usual.
    • If the vacuum pump is not running:
      • Likely a power problem or a blown breaker.
      • STOP. Report to mailing list.