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Welcome! » Transmission Electron Microscopy » Operation of the Hitachi H-600


Operation of the Hitachi H-600

The Hitachi H-600 stays under vacuum at all times. Only the Col. Switch is turned off at the end of the day or end of a session. The Col. switch turns power off and on to lenses on the column:

  1. Turn on Col. switch on right front console. See the H-600 Diagram
  2. Make sure 2 green lights are on. One on left front of console (EVAC) and the other on right front console (Camera). Both of these must be on before HV can be energized.
  3. Cool the microscope with LN2.
  4. Load specimen and evacuate specimen chamber.
  5. Insert specimen holder to first stop in microscope (stand by position).
  6. Turn on power to the voltmeter.
  7. Turn on HV (upper left console) in increments (25, 50, 75 etc.) to HV desired.
  8. Set the magnification to 4,000X
  9. Remove the Objective and Selective Area Apertures if in.
  10. Filament ImageSlowly turn up the filament current until you reach 15 volts on the voltmeter.
    --Turn the Bias control clockwise until you get emission from the tip.
    --Come to crossover with brightness controls. You should see the filament image at this point. If you don’t, adjust the Bias to read 10 Amps (one division).
    --Slowly increase the filament current until the filament image appears, you may have to adjust the crossover point using the brightness adjustment.
    --Next adjust the condenser stigmator to obtain the sharpest filament image.
    --Once you have the condenser adjusted, slowly increase the Filament current until the image of the filament just disappears.
  11. Adjust bias so that HV will read one division above KV chosen. If you choose 75 KV, then the meter should read 80.
  12. Choose a spot size by pressing 1 on the computer, then function. We normally start with a 5-micron spot.
  13. Set mag at 3000x. Check the brightness centering and condenser aperture alignment (see later section for details).
  14. Insert specimen into beam path.
  15. Center some structure- -organelle, hole, dirt- -on screen.
  16. Press CPU Reset then go to 20,000x magnification. Recenter image if necessary.
  17. Focus the image using the Z-axis control.
  18. Press "HV Modul" switch (to right of HV button).
  19. Image should move straight up and down. If it doesn’t, correct with beam BD tilt knob at bottom console. Go up in magnification and correct beam tilt if necessary. Repeat this process until you have corrected for beam tilt above the magnification you plan to work at. Turn off HV Modul Switch.
  20. Set mag control at 3,000x and insert objective aperture. The third stop is a 50 micron aperture and will be about the size of the inner screen. Center the aperture on screen.
  21. Photography (Be sure to set Counter Number for your lab if applicable).
    1. Frame point of interest and focus.
    2. Lower screen.
    3. Set illumination with brightness keys until the green light appears on photometer. We use 10-1/2 on the photometer for 2 seconds. (Dupont film).
    4. Lift right lever (shutter) all the way up and let go. It will stay up. Notice exposure flashing on CRT. When exposure is over, flashing stops and shutter can be lowered. Film automatically advances and next exposure is ready. If "error" appears on CRT, Then the camera is probably jammed. No further pictures can be taken until this is corrected. See facility staff to correct this.
    5. 30 exposures can be made from a full film cannister. The number of unexposed films is listed on CRT.
  22. Removing and Reloading Film.
    1. Turn off HV and filament current.
    2. Turn camera EVAC-Air switch to AIR. This vents the camera chamber so it can be opened. This takes about 60 seconds. Meanwhile, turn on Safelight, open film dessicator to air and turn out all room lights and panel lights.
    3. Open camera chamber door. Lift up, it swings down.
    4. Remove receiver (lower box with exposed film) and its lid.
    5. Remove film cannister (upper box). Remember that the box is upside down so hold your hand under it. Remove all loaded cassettes from camera chamber and put in camera box. Place both boxes on counter in developing area.
    6. Remove full film cannister (30 exposures) and empty receiver from dessicator and reload microscope at once. Leave receiver lid in place in the microscope but remove film cannister lid after cannister is fully inserted. Close camera chamber door and turn camera EVAV-AIR switch to EVAC.
    7. Set unexposed film mode on microprocessor to 30.
    8. Microscope is ready to use again when green camera light comes on.
  23. Developing Film:
    1. Remove film from cassettes and put in developing holder.
    2. Develop 4 min in 1:2 dilution of D-19, 68 C is optimum.
    3. Quick tap water rinse.
    4. Fix 5 min in Kodak fixer.
    5. Quick tap water rinse.
    6. Hypo clearing agent—2 min.
    7. Rinse at least 5 min in running tap water. Dry in film dryer.
  24. Loading Empty Cassette.
    1. Load film, emulsion side (not as glossy) up, making sure film is under flange on each side and tab at top. Load cassettes upside down into film cannister paying attention to direction of arrow. Put lid on cannister.
    2. Replace film cannister, spare receiver and spare film in dessicator. Put lid on dessicator. Put lever on dessicator in open position.
    3. Hold lid down with left hand fairly firmly and turn EVAC-AIR switch on pump to EVAC. Pull up on lid. If it doesn’t come off, let go. Pump until pump noise subsides. Put lever in CLOSED position and turn EVAC-AIR switch to air.
  25. Condenser Aperture Centering.
    1. For normal TEM, we use the first aperture (400 micron).
      1. With beam at crossover, insert 1st condenser aperture.
      2. Center with brightness centering knobs.
      3. Set mag control at 3,000x and spread beam with brightness button until it’s the size of the small inner-viewing screen. Center beam with aperture drives.
      4. Come to crossover and center beam with brightness centering knobs.
      5. Spread beam with brightness control and the beam should spread evenly; then comes back to center.
      6. Repeat c, d, and e if necessary until there is even spreading of the beam from crossover.
  26. Stigmatism Correction.
    This is not necessary each time you use the microscope, but if the resolution is poor, then check for a stigmatism.
    1. Put carbon hole grid in specimen holder, then into microscope.
    2. Select the highest mag that you will be using.
    3. Select the appropriate objective aperture for your use. You must always compensate at the highest mag at which you expect to work and with the objective aperture that you will be using. Each aperture has its own "personality" and must be compensated upon use.
    4. Find a round hole that you can see entirely through the binoculars at the mag you’ve chosen.
    5. Observe fresnel rings at edge of hole. You can see these best if you keep the illumination as low as possible, but yet able to see.
    6. Adjust objective stigmator (x and y) on left front console until fresnel ring is symmetrical around hole.