eTraining Introduction

Specimen Preparation

Leica Ultracut UCT

Histology

JEOL JSM-6400 SEM

Hitachi S-4700 FE-SEM

Hitachi FB-2000A FIB

Microanalysis

Veeco Dim 3000 AFM

Fluorescence Microscopes

Support

Advanced Imaging

High Resolution

1. TIP: Use a finely focused probe.    
 1.1. Use a small working distance.
 1.2. Improve signal quality by tilting the sample towards the detector.
 1.3. Use a low voltage for medium to high atomic number (Z).
 1.4. Use a higher voltage for low atomic number (Z).
2. Log onto the computer, load the sample, and turn on the microscope. See Operating Procedures under JEOL JSM-6400 SEM.    
3. Set the CL coarse to 10 using the PROBE CURRENT knob. Set the aperture (objective aperture assembly) to 4.    
4. Move the sample as close to the detector as possible by decreasing the working distance as much as possible.    
5. At high resolution, focus and stigmation are critical to quality. Adjust the stigmation and the focus. See the Focus and Magnification section of Basic Imaging under JEOL JSM-6400 SEM.    
6. Increase the magnification as much as possible while keeping the image as sharp as possible.    
7. Change the CL coarse to 14.    
8. Adjust the SE IMAGE BRIGHTNESS knob and SE IMAGE CONTRAST knob. Do not touch the focus! VIEW
9. Capture the image. NOTE: The image will appear noisy on the SEM screen, but the picture on the computer will be clear.    
10. Save the image.    

Backscatter

1. TIP: These images have the best quality when the working distance is small or the CL setting is high (4 or 5).    
 1.1. Example 1: If the WD = 39, then the CL coarse = 4-6.
 1.2. Example 2: If the WD = 15 or smaller, the CL coarse = 9-10.
2. Log onto the computer, load the sample, and turn on the microscope. See Operating Procedures under JEOL JSM-6400 SEM.    
3. Set the aperture to 1.    
4. Locate the Backscatter Detector Switch (BSE preamplifier) on the column table. Set the switch to "SL". VIEW
5. Adjust the focus and the SE IMAGE BRIGHTNESS knob and SE IMAGE CONTRAST knob. VIEW
6. Make your initial settings.    
 6.1. Control Panel 2, BE IMAGE CONTRAST knobs, set the "Coarse" knob to 4. VIEW
 6.2. Control Panel 2, BE IMAGE CONTRAST knobs, turn the "Fine" knob fully clockwise.
 6.3. Control Panel 2, BE IMAGE BRIGHTNESS knobs, turn the "Coarse" knob fully clockwise. VIEW
7. Press the IMAGE SELECTOR COMP button to display a compositional image. VIEW
8. Press the SCAN SPEED SLOW button. VIEW
9. If there is no image, that means the brightness is too high.    
 9.1. Control Panel 2, BE IMAGE BRIGHTNESS knobs, turn the "Coarse" knob counterclockwise.
 9.2. If the image is still too bright, for the BE IMAGE BRIGHTNESS knobs turn the "Coarse" knob fully clockwise again. For the BE IMAGE CONTRAST knobs, set the "Coarse" knob down one (to 3).
 9.3. Repeat steps 9.1 and 9.2 until an image appears.
 9.4. If the image is still not visible, use the CL coarse knob to lower the brightness.
10. When the image is visible, use the DISPLAY WFM button and adjust the fine BE IMAGE BRIGHTNESS knobs and BE IMAGE CONTRAST knobs. VIEW
11. Acquire and save images. See Acquiring a Digital Image in Basic Imaging under JEOL JSM-6400 SEM.    
12. When you are done backscatter imaging, return all settings to the original positions from step 6 in Backscatter.    

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