eTraining Introduction

Specimen Preparation

Leica Ultracut UCT



Hitachi S-4700 FE-SEM

Hitachi FB-2000A FIB


Veeco Dim 3000 AFM

Fluorescence Microscopes


Operating Procedures

1. Sign into the logbook. VIEW
2. Remove the dust covers. VIEW
3. Push the power button to the "ON" position. It is located behind the control console. VIEW
4. Insert the specimen block into the specimen holder and tighten. VIEW
5. Insert the specimen holder into the segment arc and tighten. VIEW
6. Set the knife into the knife block and align it with the left hand corner using the stops. Tighten. VIEW
7. Unlock the knife block. VIEW
8. Look through the microscope and shift the knife block with the knife close to the specimen surface. VIEW
9. Using the reflection of the knife blade in the specimen surface, align the knife edge with the specimen block. Lock the knife block with the lever. VIEW
10. Water is used to facilitate the making of TEM samples. Decide if water is needed.
 10.1. If using water:
  10.1.1. Unwrap the tubing from around the knife block. VIEW
  10.1.2. Loop the tubing through the roller. VIEW
  10.1.3. Fill syringe with DI water. Attach syringe to tubing. To obtain water, advance the roller. VIEW
  10.1.4. Place the refiller syringe so that the tip is inside the knife boat. Fill the knife boat with water. VIEW
  10.1.5. Adjust water amount with roller. VIEW
 10.2. If not using water, keep the tubing wrapped around the knife block.
11. Set the cutting window using the START and END buttons. VIEW
12. Select the proper cutting speed and section thickness by using the stored values in the feed menu (SEMI for approach and mirror finishes, and ULTRA for thin sectioning). Alternatively, use your own values by adjusting the - + selectors. VIEW
13. Begin sectioning.
 13.1. Auto sectioning is possible by pushing "M" in the cut menu. VIEW
 13.2. Manual sectioning can be accomplished by using the handwheel. VIEW
14. Section until the entire specimen face is cut. VIEW
15. To stop auto sectioning at any time, push "M" again.
16. If creating a mirror finish:
 16.1. Section until surface is mirror smooth, and sufficient sample is exposed from the epoxy.
 16.2. Shift the knife over to a fresh edge of the blade to complete the last few sections.
17. If creating TEM samples:
 17.1. Sections must be thinner than 100 nm. Float individual sections on water. Use an eyelash brush to separate sections from one another. VIEW
 17.2. If needed, create one section at a time using the manual handwheel.
 17.3. To lift one section from the water, place a loop around the floating section and lift straight up. The section will be suspended in a water droplet. VIEW
 17.4. To place a section on a TEM grid, lay the grid over filter paper and set the loop with the floating section over the center of the grid. The water will pass though the grid and be absorbed by the filter paper, leaving the section on the grid. VIEW
 17.5. Repeat as many times as necessary to create the perfect TEM sample. VIEW
18. When sectioning is complete, stop the motor (by pressing "M").
19. Unlock the knife block with the lever and retract from the specimen.
20. Unlock the specimen holder from the segment arc.
21. Remove the specimen block from the holder.
22. If using water:
 22.1. Remove the refiller syringe from the knife boat.
 22.2. Dispose of the knife boat water.
 22.3. Use an air can to clean the boat.
 22.4. Remove the tubing from the roller.
 22.5. Dispose of the water from the tubing and syringe.
 22.6. Wrap the tubing around the knife block, and replace the refiller syringe onto its magnetic base.
23. Clean the knife blade. Pass a Styrofoam stick across the blade to remove any debris. VIEW
24. Turn off the power and replace the dust covers.

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