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Microanalysis

WDS Quantitative Analysis Procedure

1. Prepare Sample | 2. Start Up | 3. Select and Set Stage Positions for Sample & Standard | 4. Aligning OM with Electron Beam Position | 5. Setting up the Analysis Parameters | 6. Calibrating Standards | 7. Analysis Template Setup | 8. Set & Store Sample Stage Positions | 9. Perform Analysis | 10. Review Results | 11. Shut Down

1. Prepare Sample

1.1. The sample and standard should be placed in a two-position specimen holder. Keep track of which one is which. The standard should be properly positioned so that it is aligned with its paper map. NOTE: You need to measure every element that may be present in the sample. If you do not, you will get numerous errors in the WDS results. Perform an EDS analysis first if you do not know all the element constituents. VIEW
1.2. Apply carbon conductive paint over the standard ring edge and the ring holder edge. VIEW
1.3. Repeat twice for both the sample and the standard (There should be 8 drops of paint total.) Do not cover the standards or your sample with the paint! VIEW

2. Start Up

dQant32 Icon2.1. Log onto computer and load dQant32 software. VIEW
ESP Icons2.2 Hit [E][S][P] icons in the dQant32 program (element, standard, points).    
2.3. Load the sample and start up the SEM. See Operating Procedures under JEOL JSM-6400 SEM.    
2.4. kV = 20, WD = 39, Tilt = 0.    
2.5. Adjust using the SE IMAGE BRIGHTNESS knob, SE IMAGE CONTRAST knob and MAGNIFICATION knob. VIEW
2.6. Turn on the picoammeter. VIEW
2.7. Push the blue PCD (Probe Current detector) button (lights up) on the control console to insert the probe current detector into the beam path. VIEW
2.8. Press the [EOS] key on SEMs keyboard to launch a menu on the right SEM screen. Set beam current to 20 nA on the picoammeter using the CL. VIEW
2.9. Set aperture (objective aperture assembly) to 1. VIEW
2.10. Set CL coarse = 7, CL fine = ~55. VIEW
2.11. Readjust CL coarse until the picoammeter reads 20 nA. When finished, turn off blue PCD button (light goes off). VIEW
2.12. Over the first ½ hour, the beam current will drop. To correct, adjust the CL fine to a smaller number. VIEW
2.13. Flip the PCD switch from internal to external. VIEW
2.14. Use the upper left black button on the joystick to move the PCD in and out of the beam path. Only use this button to toggle between in and out from now on. VIEW

3. Select and Set Stage Positions for Sample & Standard

3.1. Crank the EDS detector to the "in" position. See the NOTE in step 1.1 about elemental analysis before proceeding. VIEW
3.2. In the "Standard Table: Master", locate the standards you will use. To highlight these, hold [Ctrl] and select your required standards, then release [Ctrl] and right click the mouse. Hit "Show Selected Records" to show only these selected standards in the Standard Table window. VIEW
3.3. Use your standard map to locate these standards in the SEM. (As an aid in locating the standard, from the "dSspec" window, select menu "Stage > Specimen Holder Map".) Click on the stage map graphic where you want to move, then hold [Ctrl] and left click on the mouse at the same time. VIEW
3.4. Highlight your standard name in the "Standard Table: Master". Click the "Set" button at the bottom of the table. A "Setup Standard" dialogue will appear. Click "Store".    
3.5. Set and save the stage position for standards and Benoite (MAC), or Willemite (ARL). These are the fluorescing samples needed later. Refer to the STD map. Use the EDS to ensure that the elements agree with the map. VIEW
Spectrum Acquisition IconErasure Icon3.5.1. If in doubt about the standard's composition, choose Spirit software, click "erase", and "acquire".
3.5.2. Adjust the beam current so EDS dead time = 25-30%.
Stop Spectrum Icon3.5.3. Stop acquiring, label peaks, go back to dQant32 screen. Go to the Standard Table and highlight your standard name. Click "set" and "store". Repeat above steps until all standards positions have been updated.
3.6. Select "Point Table" in dQant32 screen. Select menu item "Tables > Add New Table". Name it.    
3.7. Move to the sample using "Specimen Holder Map". Save the stage position for your general work area in the Point Table.    
3.7.1. Place the cursor in the first line of the new Point Table.
3.7.2. Click "set" and "store".
3.7.3. Repeat for all samples.
3.8. NOTE: The software will support a Quantitative Line Transect. See support staff for instructions.    

4. Aligning Optical Microscope (OM) with Electron Beam Position

4.1. Flip the PCD to "in" and check for changes in beam current. Adjust the fine CL until 20 nA of beam current is displayed on the picoammeter.    
4.2. After the stage has moved, flip the PCD back to "out". If the screen is black, the PCD is "in", and the beam current will drop when its toggled.    
4.3. Focus in on a standard that is fluorescent. (Select "Standard Table" on dQant32 screen, select either Benoite or Willemite line, and click "Get".)    
4.4. Carefully focus the image on the SEM using the objective lens.    
4.5. Tilt the Optical Microscope (OM) down by grasping and pulling. VIEW
4.6. Move the OM to the "in" position. You should be focused in on the fluorescing sample. VIEW
4.6.1. NOTE: If at any time your standard is not visible, or you can not find it, the OM must be moved back out.
4.6.2. Magnification must be 50x or greater.
4.7. Press [PF6] to turn off the SEM scan mode. VIEW
4.8. Make sure the PCD button is "out" on the joystick control.    
4.9. Locate the illumination from the beam through the OM. VIEW
4.9.1. Take the cap off. VIEW
4.9.2. Turn the LCD screen on. VIEW
4.9.3. Pull the knob out (on left side). When the knob is out, you can view through the objective lens. When the knob is in, you can view the image on the LCD instead. VIEW
4.9.4. Adjust OM using the locknut until the beam is in the center of the optical field.
4.9.5. Use the fine Z-axis The fine Z-axis stage translation is used to make fine changes in the distance between the specimen surface and the objective lens. SEM stage. control on the SEM to focus the illumination dot in the OM.
4.9.6. Push the knob back in. The LCD screen should show the illuminated dot. VIEW
4.9.7. Push the green [ON] button on the illumination console. The dot should be almost centered on the screen.
4.9.8. Refocus the image features (not the dot) with the Z control. VIEW
4.9.9. Use the objective lens (PROBE CURRENT knob on control panel 1) knob to produce the smallest dot possible. VIEW
4.10. Pull the knob out (on the left side). VIEW
4.11. Move the OM crosshairs to coincide with the beam position on the fluorescing sample using the two knobs on the eyepiece VIEW
4.12. Flip the objective lens polarity switch on the control console twice (this removes hysteresis from the objective lens), then refocus with the objective lens and adjust crosshairs. Do not touch the objective lens setting after this point and do not change the crosshair position. VIEW
4.13. Toggle the PCD in. Double check the beam current and adjust it back to 20 nA if necessary.    

5. Setting up the Analysis Parameters

5.1. In the dQant32 program, select the Element Table. Select menu "Tables > Add New Table" and name it.    
5.2. Select menu "Element > Open Second Window". "Element Table2: 20KV" opens. VIEW
5.3. Select "Element Table2: 20KV". Select an element by clicking in the first column. Drag that element into the "Your named Element Table". Repeat for all desired elements.    
5.4. Select the first element in "Your named Element Table". Select "Standard Table: Master" and choose the standard name for that element. Click "Copy To Ele", and the standard name will be inserted into "Your named Element Table". Repeat for all elements in "Your named Element Table".    
5.5. In "Your named Element Table" select an element. Click "Get Std" in the buttons below the element names. The stage will move to the updated coordinates of that elements standard (provided you saved the coordinates previously). NOTE: Do not move the joystick while the stage is in motion! See support staff if stage and motors have lost communication and the joystick no longer moves the stage manually.    
5.6. If your Gaussian peaks are not present, follow the steps in the Troubleshooting topic in JEOL JSM-6400 SEM.    

6. Calibrating Standards

6.1. Go to "Your named Element Table". Select the desired element and click "Get Std". Check the OM focus and adjust with fine Z-axis control.    
6.2. Hit "CSearch" (coarse search) button. This drives the spectrometer to a within a few millimeters of the peak location and opens the Probe Current detector.    
6.3. In the peak search window you will see a representation of the counts.    
6.4. You should see a Gaussian shaped peak. If not, follow the steps in the Troubleshooting topic in JEOL JSM-6400 SEM. If that does not work, contact support staff.    
6.5. You may see error boxes appear. Hit "continue" until the error boxes go away.    
6.6. Coarse search is already in place, so now go to fine search by clicking the "FSearch" button.    
6.7. Now a higher resolution Gaussian peak is collected.    
6.8. Click "PkCalib". When the stage moves, its called a "jog" to relieve gear lash.    
6.9. "Focus Standard" dialog appears. Hit close.    
6.10. The "dSspec" window appears. Counts will be displayed. You will hear the detector moving. When the calibration is finished the new standard values will be stored, and displayed in "Your named Element Table".    
6.11. Repeat three times by hitting "PkCalib" 3 times.    
6.11.1. Each time you hit "PkCalib", hit "close" on the "Focus Standard" dialogue afterwards.
6.11.2. Watch the peaks collect in "dSspec".
6.12. Go to "Element"on the dQant32 menu and select "Average Calibration File…". The bottom entries are the most recent.    
6.13. Make sure the last three calibrations are similar under "net peak".    
6.14. Highlight those three rows (hold "Ctrl" while selecting). Hit "Apply Average" then "Close".    
6.15. Repeat steps 6.1 - 6.14 for each element that you want to measure.    

7. Analysis Template Setup

7.1. Now set up the quant file. Go to the "Quant" menu and choose "Setup…".    
7.2. Hit "New Setup" button.    
7.3. In the "Setup File" field, replace the "New File" path name with your new file name.    
7.4. In the "Title" field add the title of the project.    
7.5. Input 20 to the keV field.    
7.6. In the "Dataset Folder" field, highlight the previous file name at the end of the path and replace it with your own file name. A dialog will tell you that the folder does not exist. Click "Yes" to create it. Alternatively, you can open "My Computer" on the desktop and create a folder, then type that path instead into the "Dataset Folder" field.    
7.7. Enter a unique name in the "Dataset File" field.    
7.8. Click "From Ele Table". It imports data from "Your named Element Table" to the "Quant Setup" table. The "Reference" column in "Quant Setup" should match the "Name" column in "Your named Element Table". Hit "Close". Click "Yes" to save changes made to the Quant Setup.    

8. Set & Store Sample Stage Positions

8.1. Go back to dQant32 "Your named Point Table", highlight the sample stage position, and click "get" (stage moves automatically). This moves to the position on the sample that you previously saved.    
8.2. Press [PF7] to turn on SEM scan mode. VIEW
8.3. Check that the PCD is out.    
8.4. Lower the magnification.    
8.5. Adjust contrast and brightness.    
8.6. Reposition the sample to your desired analysis area using the stage control. Store the position in dQant32 points table. Be sure to write down which numbers correspond to which analysis points. (A line scan can be set up using this points table.)    
8.6.1. Single point: Move stage to the desired location. All positioning will be done with the joystick while viewing the OM screen.

(If necessary, in the dSspec window, under the "Joystick" menu, select "Orientation Reversed" for easier control. Control will then be reversed in the SEM screen.)

Refocus with fine Z-axis. In "Your named Point Table", select the corresponding point number. Hit "Set". "Setup Point" dialog appears. Hit "4 - Store" and "Close".
8.6.2. Multiple points: Make a note of stage coordinates for your other specimens. Repeat step 8.6.1 above multiple times, once for each desired location.
8.6.3. Line of points: Repeat step 8.6.1. Reposition the stage to the location at the far end point for your line. In "Your named Point Table", select the second point number. Hit "Set". "Setup Point" dialog appears. Hit "4 - Store" and "Close". Then click "Calc. Line". The "Line Scan" dialog appears. Choose the number of points in your line OR the distance between points. NOTE: If you require evenly-measured spacings between points, it is easiest to move only in the X or Y directions.
8.7. Press [PF6], view the sample through the OM, and focus with the fine Z-axis. Toggle the PCD in. Adjust beam current to 20 nA using the condenser lens for fine control. Press [PF7].    

9. Perform Analysis

9.1. Hit "Macro", "Run…" and open "QUANT.MAC". See the NOTE in step 1.1 about elemental analysis before proceeding.    
9.2. Go to "dSspec" window and observe spectrometer activity. When measurements are complete, activity ceases and a sound is issued.    
9.3. Results (Measure Table) appear in the "Output" window. (They will also be stored in an MS Excel file for retrieval.)    
9.4. Evaluate the results. That is, does the weight percent element = 100% +/- 2.5%? If so, conclude that the calibration is correct. Continue with your measurements. If the calibration is not correct:    
9.4.1. OM focus is bad.
9.4.2. OR you chose the wrong standards for your calibration. (Example: Using Si metal as a standard for SiO2 measurements OR assuming your sample is homogeneous when it is not.) Use a standard of material similar to the specimen.

10. Review Results

10.1. The results will be readable with Microsoft Excel. Locate it in the dataset folder. Do Quant, Setup, and view the path to "Dataset Folder". Navigate to that folder on the computer.    
10.2. Alternatively, do "Quant > View Dataset".    

11. Shut Down

11.1. If you are done and want to quit, flip PCD switch back to internal.    
11.2. Press [PF7]. VIEW
11.3. Retract OM, replace cap, and tilt arm up.    
11.4. Reset aperture to position 4. Adjust contrast and brightness.    
11.5. CL = 10.    
11.6. Stage home position: dSspec > Stage > Home Position.    
11.7. EDS out. VIEW
11.8. Save and close all programs. Unload specimen.    
11.9. Shut off SEM as usual. See Shut Down under JEOL JSM-6400 SEM.    

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