Using the JEM-1010 Transmission Electron Microscope for thin resin sections of biological material

Inserting the sample:

1. Ensure the HT and filament are turned off
2. Pull the specimen holder straight out until it won’t go any further, then gently turn it anticlockwise until it stops
3. Very gently pull the specimen holder out and rest it on the stand. Make sure you never handle past the O-ring area towards the sample end
4. Carefully flick up the lever at the sample end with forceps
5. Load the grids into the specimen holder (dull side up; i.e. sample side up) and gently lock in place with lever
6. With a flat palm, push the specimen holder in softly so that the guide pin (on left) fits into slot. The green light at right will come on.
7. After the vacuum has pumped through one cycle the green light will go off.
8. Turn the specimen holder clockwise a quarter of a turn. It will stop. Do not force it further.
9. Guide the specimen holder in gently.

Initial viewing of an image:

1. Check HT i.e. if it needs changing. If so then:
   • Turn OFF HT (never change HT with HT on)
   • Type ‘htset 80’ <enter> (for 80 kV)
   • Alternatively, ‘htset 100’ <enter> (for 100kV)
2. Turn HT ON (it will light up when pressed)
3. Wait for beam to stabilize (53 µA for 80 kV)
4. Turn filament ON

5. Chamber screen should now be filled with green light.
6. If there isn't any light, ensure the camera is out (press red light on camera control); check that the magnification is not extremely high; or possibly a grid bar is blocking the beam.
7. To find your sample on the grid, press LOW MAG and move out the objective aperture by turning the silver lever to the right and you will see the full grid.
8. Using the big white wheels (left and right of column) move your sample around, find the area you want and bring it to the center on the screen.
9. Move the objective aperture (on column below sample port region) in by turning the silver lever to the left.
10. Press MAG 1 and spread the beam (brightness knob on left hand side console).
11. Realign the objective aperture.

Things to check:

Objective aperture (located on column below sample port region)

1. At around 5000x mag. adjust the brightness knob (left hand console) to create a small bright circle.
2. Press DIFF (right hand console) and you should see a bright circle in the centre of a dull circle.
3. If not, center using the objective aperture two knobs.
4. Press MAG 1 and spread the beam.

Condenser aperture (the top adjustable aperture on the column)

1. Should be set at largest.
2. To change, turn large condenser knob with dots on it to appropriate setting.
3. If not aligned then the beam swings across the screen when brightness is changed.
4. To correct, adjust the brightness knob (left hand console) to create a small bright circle.
5. Centre the beam with SHIFT X and Y (left and right hand consoles) then spread beam clockwise until it is nearly the same diameter as the screen by turning the brightness knob (left hand console).
6. If off center then use the condenser aperture knobs to correct position on screen.

Grid height and Wobbler

1. With camera in, at around 50,000 to 80,000 mag undertake the following:
2. Check to see that the objective lens current is correct (page 6 on microscope computer screen). i.e. 3.85 at 80 kV. Use focus knob on x16 (right hand console) to adjust the number.
3. Focus using the z-knob (small knob directly under the specimen holder).
4. Take out the camera.
5. Set mag, to 3000 to 5000x.
6. Using brightness knob (left hand console) bring beam to a small bright circle.
7. Press X Wobbler (right hand console).
8. Use Wobbler knobs under the Perspex cover (right hand console) to bring both spots together on screen.
9. Turn off Wobbler.
10. Spread beam (brightness knob – left hand console).

Spot size

1. This should be around 1 (large) if the sample is stable or 5 (small) if drifting.
2. To change, bring beam to a bright circle using the brightness knob (left hand console).
3. Use the flat buttons (left hand console) to choose a spot size.
4. If the beam moves across the screen then re-center using SHIFT X and Y (left and right hand consoles).

Hints for objective astigmatism adjustment at magnifications greater than 80,000x

1. Press Obj Stig (left hand console) button and use Def X and Y (x16 if course adjustment required). [Def = deflector]
2. Bring microscope to neutral astigmatism position by ensuring both green lights on Def X and Def Y are lit.
   1. Whilst looking at image and using Def X, Def Y and focus (one at a time) create best image possible.
   2. Whilst looking at the RTFFT (Real Time Fast Fourier Transformation) panel on computer screen and using Def X, Def Y and focus, create the largest and roundest circle possible.

Changing objective aperture sizes

1. Camera OFF and out.
2. With the sample IN, at around 5000x mag, adjust the brightness knob (left hand console) to create a small and bright circle.
3. Press DIFF (right hand console). You should see a bright circle in the center of a dull circle.
4. Change aperture using the spots on the side as a guide to size.
5. Ensure the bright light is in the center of the dull light. If not, center using the black objective aperture knobs (at front and right of aperture knob).
6. Press MAG 1.
7. Spread the beam (brightness: knob on left hand console).

End of session

1. Camera OFF and out of chamber.
2. Condenser aperture at largest (1).
3. Magnification under 15,000x.
4. Spot size at 1.
5. Turn filament and HT OFF.
6. Remove grids and replace sample holder into microscope.
7. Close the digital capture system (ITEM), turn off computer screens, replace over on screen port and turn HT on.

Last client of the day:

1. Turn HT off.
2. Scroll brightness on screen down.