COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK COLUMBIA NANO INITIATIVE / CENTER FOR INTEGRATED SCIENCE AND ENGINEERING

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Zeiss Sigma VP SEM Standard Operating Procedure

These instructions are intended for reference only, and will not replace the thorough training required for proper system operation. Contact an electron microscopy staff member with questions or to report a system problem. This SOP was written with the help of Scott Trocchia.

COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK COLUMBIA NANO INITIATIVE / CENTER FOR INTEGRATED SCIENCE AND ENGINEERING

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1. BADGER IN: Enable the tool in Badger.

2. SOFTWARE: The user interface is called Smart SEM (can be found on the left screen). Check the EM Server (on the right screen) to make sure there are no error messages. If there are please let the staff or superusers know.

COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK COLUMBIA NANO INITIATIVE / CENTER FOR INTEGRATED SCIENCE AND ENGINEERING

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3. MOUNTING SAMPLES: Put on pair of gloves and mount your samples on the stubs using carbon tape. Use as little tape as possible.

4. VENTING: Make sure EHT is off. Click VENT button on the toolbar, then wait approximately 1 minute. The Vac Status label in the data zone (white bar at bottom of viewing area) should read “At Air”.

COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK COLUMBIA NANO INITIATIVE / CENTER FOR INTEGRATED SCIENCE AND ENGINEERING

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5. OPENING CHAMBER: Use the handles to open the door. Take the carousel (sample holder) out and close the door completely. Place sample stubs into designated holes on the carousel and screw to tighten. Open chamber, put holder back into SEM, and close the door.

6. PUMPING: Click the PUMP button on the toolbar. Wait for (Vac status = Ready, System Vacuum ~ 2.0e-5) in the data zone.

COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK COLUMBIA NANO INITIATIVE / CENTER FOR INTEGRATED SCIENCE AND ENGINEERING

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7. SAMPLE ADJUSTMENTS: Adjust specimen height, if necessary (default: 3 mm). Move to desired sample slot by using X/Y/R joystick (hardware) or double-clicking carousel graphic (software). Adjust height of stage using Z joystick, noting that the Z-movement automatically stops at the blue line drawn in the Stage Navigation Bar.

COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK COLUMBIA NANO INITIATIVE / CENTER FOR INTEGRATED SCIENCE AND ENGINEERING

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8. EHT: Set EHT (0 to 20 kV) by double-clicking the word in the data zone. Turn EHT ON by pressing corresponding button on toolbar. Switch to imaging mode by clicking the TV button on the toolbar.

COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK COLUMBIA NANO INITIATIVE / CENTER FOR INTEGRATED SCIENCE AND ENGINEERING

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9. INITIAL IMAGE ADJUSTMENTS: Use magnification knob on keyboard panel to set magnification to a minimum. Adjust brightness and contrast using knobs on keyboard panel in order to see relevant features on your sample(s). Aim for a starting working distance close to 4 mm. Make sure that the detector written in the data zone (“Signal A = ”) label matches the detector you are using. InLens corresponds to the primary detector.

Magnification

COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK COLUMBIA NANO INITIATIVE / CENTER FOR INTEGRATED SCIENCE AND ENGINEERING

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10. FOCUSING IMAGE: Identify region of sample to image. Zoom in as much as possible using magnification knob. Turn the Focus knob on the keyboard panel to focus as much as possible. Use the Stigmator X & Y knobs on keyboard to minimize image stretch/distortion upon moving in and out of focus.

COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK COLUMBIA NANO INITIATIVE / CENTER FOR INTEGRATED SCIENCE AND ENGINEERING

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11. APERTURE ALIGNMENTS: Click the Focus Wobble checkbox. Use wobble amplitude of 10-30%. View the Aperture Align graph, and use the Aperture X & Y knobs on keyboard panel to minimize image oscillations. If you can, it is recommended to use >10,000X magnification for aperture adjustments.

12. CORRECTING ASTIGMATISM: Un-check Focus Wobble checkbox. View Stigmation graph and optimize the image distortion using the Stigmator X & Y knobs and the Focus knob.

COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK COLUMBIA NANO INITIATIVE / CENTER FOR INTEGRATED SCIENCE AND ENGINEERING

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13. FURTHER IMAGE ADJUSTMENTS: Iterate steps 9 through 12 until you are satisfied with the image quality you see on the screen.

14. SAVING IMAGES: Choose Scan Speed 5/10. Freeze (red dot in bottom right corner of data zone signifies ready to save). Save (TIF, JPG, BMP, all with or without Annotation). Make sure to navigate to your user directory before confirming the saving operation.

15. FINISHING UP – SOFTWARE: Reset Scan Speed to 1. Go to low (< 100X) magnification. Turn EHT OFF.

COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK COLUMBIA NANO INITIATIVE / CENTER FOR INTEGRATED SCIENCE AND ENGINEERING

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16. FINISHING UP – HARDWARE: Move stage height down to safe zone (30 mm). Move to sample slot #9 (geometric center of carousel). Rotate and tilt stage back to 0 degrees.

17. VENTING: VENT. Remove sample(s). Place carousel/holder back inside chamber.

18. PUMPING: PUMP down to (Vac Status = Ready, System Vacuum < 1e-4).

19. CLEANING UP: Clean and tidy desk and SEM table. Report to superusers if any issue occurred during your reservation time.

COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK COLUMBIA NANO INITIATIVE / CENTER FOR INTEGRATED SCIENCE AND ENGINEERING

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20. BADGER OUT: Disable the device in badger and log out.