Using the Hitachi 3400-N VP-SEM Opening the Chamber to Load Specimens (This may also be done later using the software) 1. Click the AIR button on the front of the machine: 2. Wait a few minutes until you hear the machine beep 3. Always wear gloves when handling anything that goes in the chamber-stubs, specimens, sample holders etc. 4. Measure your sample s diameter and height. EVAC AIR 5. For multi-holder, insert with sample #1 facing blue pen mark in chamber 6. Close the chamber and hold it shut with your hand. Press EVAC button on front of machine. Let go of chamber door when vacuum engages it. Starting PC-SEM 7. Click the PC-SEM icon on the desktop. There is no password. 8. When the PC-SEM finishes booting, you will see the Specimen Dimensions window at right. Select the correct Diameter/Size and Height for the sample. Click OK. 9. On the rights side of the screen are 4 overlapping control panels-condition, Image, Utility and Stage. 10. Go to Condition and ensure you are on SEM mode and not VP-SEM. 11. Go to the Image tab and choose SEM or BSE mode as required. 12. Go to the Stage tab and set the desired stage height (z-position). 10 mm is a good starting point* Click GO. 13. Wait for specimen to reach desired height. 14. (OPTIONAL) If you are using the multi-holder, click the Memory button in the lower right corner of the stage tab. 6 positions are programmed in the pop-up window. Choose Page 1, then choose the desired sample position and click Move to go to that sample. For Multiholder Choose Page 1, select position and hit move * See staff for setting recommendations
5. Magnification Beam Settings Settings Pop-Up Window Tabs: Condition Image Utility Stage Details Button Setting up Conditions 1. On the Image Tab, make sure you are on SE and not BSE mode. 2. On the Conditions tab, ensure that you are on SEM and not VP-SEM mode (white arrow above) or you will have trouble focusing. 3. Click the Details button or double click on the Beam Settings window to open the settings window. (See screen shot above) 4. In the settings pop-up window (left), select the appropriate accelerating voltage* for your specimen and set probe current* RECORD YOUR SETTINGS HERE: Mechanical Aperture= KV= Probe Current= Stage Height= 5. Turn magnification to 100x using button onscreen or manual dial. 6. With the chamber properly evacuated and the accelerating voltage set, switch on the electron beam by clicking the ON button in the upper left. The HV progress window will open. 7. Click the AFS button to saturate the filament. It should deliver an emission current above 70uA. 8. Click the ABCC button to set Auto Contrast and Brightness. 9. Use the Fast or Slow1 scan speed to adjust focus. Increase magnification slowly to desired level while adjusting contrast, brightness and focus as you go. 10. Once you see an image, turn the mechanical aperture to the correct level-typically 1-2 for low mag and 3-4 for high. Alignments must be done whenever changing the KV, probe current, or aperture size. Get help or refer to Align the Instrument Section of this document.
11. Focusing your sample is a multi step process. Start by using fine focus to get best possible image. Then use stigmator X turn one way and then the other to find best possible image. Repeat using stigmator Y. Adjust fine focus again. Repeat this 2-3 times until you get the best possible image. 12. Adjust contrast and brightness for proper dynamic range. 13. Click the desired scan speed TV or Fast to scan your specimen and locate areas of interest; Slow3/5 to preview a high resolution image prior to capture. Capture an Image 1. In Slow3/5 scan mode click the M/2560 button, the Capturing Image progress window will open. 2. The Captured Image window will open in the lower part of the monitor as a thumbnail image. 3. The Image window will display CAPT in the upper left corner. 4. To return to live viewing, click RUN again to reactivate the detector. Saving Images 1. Select the thumbnails of those images you wish to save-they will be highlighted yellow. 2. Embed into image will burn in annotations associated with image. (To change what information is displayed on your image, go to Set-up Image Display Record) 3. Click the Save button this will open a dialogue box. 4. Click Select to find your folder first. You have to hit save twice after naming the file. 5. DO NOT KEEP > 10-15 images open, or the software may freeze up. Save and then delete them. Changing Specimens or Ending your Session 1. Click the HV OFF button in the upper left. 2. Click the AIR button in the upper right (Specimen Dimensions window will open). Select 15mm and Standard click OK. The chamber will open in about 90 seconds. 3. Using gloves or forceps, remove your specimen and replace the holder.
4. Close the Chamber door and click the EVAC button. 5. When the system has fully evacuated, close the PC-SEM Program. Leave computer on. Using Dropbox to Transfer Images 1. Get a staff member to help you setup your shared Dropbox folder. 2. At the central workstation in the lab, follow the posted directions. 3. Open a second window of My Computer and locate your image files onthe mapped drive of the SEM computer. 4. Transfer files to your Dropbox folder. 5. Keep the email link for future use. Align the Instrument: If you change the mechanical aperture, the accelerating voltage or the probe current, specific alignments of the instrument must be performed. Mechanical Alignment: -At the desire magnification, first turn the X-Align knob at the end of the aperture control until the beam reaches its brightest. -Hit ABCC to readjust ritness. -Repeat using Y-Align knob on the side of the column. You must now perform the following alignments: Aperture Alignment: -Begin with a magnification of 1000x 5000x, then repeat a second time at twice the desired magnification. -Apply the KV and probe current settings desired for operation -Perform normal focus and stigmation operations without the align window open -Open the alignment window -Select Aperture Align -Move stigmation knobs to minimize image movement -Close alignment window -Repeat at 2 x desired magnification for best results. NOTE: If focus changes substantially during routine operation, try the degauss Button in setup, then try an aperture alignment if that doesn t fix the problem.
Beam Alignment: -Select Beam Align Tilt -Turn X and then Y stigmation knobs until maximum brightness is achieved -Repeat on Beam Align Shift Stigma Alignment: -Set magnification between 1000x and 5000x -Find a noticeable feature on the image with some fine details -Apply the KV and probe current settings desired for operation -Perform normal focus and stigma alignments -Degauss Image before beginning for best results -1 st Step: Stigma Align X: Adjust to minimize rocking movement in the X- direction -2 nd Step: Stigma Align Y: Adjust to minimize rocking movement in the Y- direction -3 rd Step: repeat as needed until image just appears to move in and out towards you, not in X or Y directions. -Close alignment window. -Repeat Degauss step, then repeat procedure at 2x desired magnification Using Variable Pressure Mode This mode allows you to view uncoated, non-conductive specimens by suppressing charging. 1. You will need a higher KV to use this mode, usually 10-20KV with WD = 5mm.. 2. Set up first using a control sample on regular SE mode. Focus, stigmate and align the beam. 3. Turn KV Off. 4. Under Conditions tab, select VP-SEM. 5. Set vacuum to 30-50 (lower is more efficient) 6. Evacuate the chamber and Switch to the test sample. 7. Turn KV back ON. You will need to focus on Slow 1 or Slow 2 mode-fast is too noisy. 8. Under VP-SEM DETAILS, adjust light setting to COMP. 9. You may need to adjust your camera GAIN setting to 3 or 4.