eTraining Introduction

Specimen Preparation

Leica Ultracut UCT

Histology

JEOL JSM-6400 SEM

Hitachi S-4700 FE-SEM

JEOL JEM-4000FX TEM

Microanalysis

Veeco Dim 3000 AFM

Fluorescence Microscopes

Support

Advanced Imaging

Scanning Transmission Electron Microscopy (STEM)

1. Preliminary | 2. Initial Settings | 3. Column Alignment | 4. Scanning Image Observation | 5. Acquiring a Digital Image | 6. Acquiring X-Ray Maps

1. Preliminary

Before beginning, perform all the steps required to activate and operate the TEM from Specimen Preparation through the Basic Imaging procedures. These are:

Specimen Preparation

Operating Procedures, including Initial Alignment

Imaging Techniques > Basic Imaging

under JEOL JEM-4000FX TEM.

2. Initial Settings

2.1. Adjust the DISPLAY 1 BRIGHTNESS and CONTRAST to the 3 o'clock position. VIEW
2.2. Adjust both the SEI and TEI BRIGHTNESS to the 3 o'clock position. VIEW
2.3. Adjust both the SEI and TEI CONTRAST to the 3 o'clock position. VIEW
2.4. Depress the SEI and TEI IMAGE POLARITY switch to "+". VIEW

3. Column Alignment

3.1 Depress the SM button.

NOTE: This is a 3-position switch that toggles between STEM alignment mode (brightly lit), STEM image observation mode (brightly lit), and conventional TEM mode (dimly lit).		 VIEW

3.2. Depress the SM button once to engage stem alignment mode.

    
3.3. Set to Page 4 by engaging the PAGE button in the right drawer. VIEW
3.4. Maximize the COND 3 value with the BRIGHTNESS knob. VIEW
3.5. Turn the OBJ FOCUS knob to obtain a caustic spot. VIEW
3.6. Engage the DEFLECTOR, BRIT TILT button on the left panel, and center the caustic spot with the DEF X, Y knobs on the main panel VIEW
3.7. Adjust the OBJ FOCUS knob to the proper voltage for the accelerating voltage from the table. See Heating the Filament under Operating Procedures in JEOL JEM-4000FX TEM.

    
3.8. Focus the illumination to a spot using the BRIGHTNESS knob. VIEW
3.9. Center the illumination using the SHIFT X,Y knobs on main panel. VIEW

3.10. Repeat steps 3-8 until the caustic spot and illumination spot remain in the screen center.

    

4. Scanning Image Observation

4.1 Depress the SM button until the scanned image appears on the CRTs. VIEW
4.2 Depress the SEI button adjacent to the DISPLAY 1 (left hand) in the display unit. VIEW
4.3 Depress the NOR/D-MAG button until a full scan frame is present on DISPLAY 1. The button toggles between D-MAG, a reduced raster frame and a normal or full screen scan frame. VIEW
4.4 Depress the DET button to engage the SEI detector. VIEW
4.5 Adjust the SEI CONTRAST and BRIGHTNESS to obtain an image on display. The same image may appear on DISPLAY 2 depending on the position of the button described in step 4.2. VIEW

4.6 Astigmatism, or directional stretching of the image as you focus and defocus the image may be corrected with the STIGMATOR X and Y knobs on the STEM Control Unit. The steps are as follows:

    
4.6.1 Focus the image with the OBJ FOCUS knob on Control Panel Right in such a way as to minimize the stretching. The image will appear of out focus. VIEW
4.6.2 "Focus" the image with either of the STIGMATOR X or Y knobs. Repeat with the other stigmator knob. VIEW
4.6.3 Refocus the image with the OBJ FOCUS knob.
4.6.4 Repeat steps 4.6.1-4.6.3 until the stretching is eliminated during focusing or defocusing.
4.7 Depress the WFM on the display unit control panel. Six horizontal, stationary lines will appear on the CRT. VIEW
4.8 Use the SEI BRIGHTNESS and CONTRAST knobs to control the moving line. The base of the signal should be at the second line from the bottom and the highest signal should be just above the top of the line. VIEW
4.9 Disengage the WFM button to return to an image. VIEW

5. Acquiring a Digital Image

NOTE: Detailed manufacturer's instructions can be found in the online in v 1.6.0 Basic Operating Principles under 4pi Spectral Engine Resource Manual and Revolution User Guide in the support pages for 4pi Analysis, Inc. See Disclaimers in Support.

Revolution Toolbar

Revolution Toolbar

The software and icons refer to the 4pi Revolution® software for Windows.

5.1 Select Mode > SEM Image. VIEW
5.2 Select the Setup icon and configure the SEM Image Setup dialog as: VIEW
5.2.1 Enable Channel 1 and 2.
5.2.2 Check Frame Average and 20 frames OR uncheck Frame Average and set Channel 1 Dwell to 200.

NOTE: The Dwell = 200 option produces a single pass, high-resolution image.
5.2.3 Check Line Sync.
5.2.4 Format = TIFF-PC.
5.2.5 OK
Preview Icon5.3 Using the Preview icon, make fine adjustments to the SEI CONTRAST and BRIGHTNESS on the STEM Control Unit to appropriate levels. VIEW
5.4 Select the pixel density for your image. VIEW
Acquire Icon5.5 Select Acquire and enter the STEM magnification. VIEW
5.6 The digital SEM image will be displayed. VIEW

5.7 Save the image in your directory in the "User files" folder.

    

6. Acquiring X-Ray Maps

See EDS X-Ray Mapping under Microanalysis in JEOL JSM-6400 SEM.

NOTE: Detailed manufacturer's instructions can be found in the online in v 1.6.0 EDX Image Mode under 4pi Spectral Engine Resource Manual and Revolution User Guide in the support pages for 4pi Analysis, Inc. See Disclaimers in Support.

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