ZEISS 10A TRANSMISSION ELECTRON MICROSCOPE

OPERATING INSTRUCTIONS

Usual settings for operation

• Condenser 2 aperture: any position (do not change)
• Objective aperture: multihole aperture disk (usually position 3)
• Projection 1 aperture: out
• Condenser 1 lens control: 2
• Beam current: just below 3 o'clock position
• Accelerating voltage: 60 or 80 kV

To turn On Microscope

• Press I/O button. (Alarm will sount briefly)
• Wait for green lightening light to appear (approximately 20 min).

  If alarm warning that water sustem is malfunctioning sounds druing operation of the microscope, turn off microscope immediately.

To Insert or Remove Specimen

• Make sure filament and high voltage are off.
• Pull out specimen rod to arrest position, turn to stop in milled-out ring and pull out until it engages in stop position.
• Turn large knurled wheel fully counter-clockwise.
• Screw key into slide, remove slide by gently pulling on key, and place slide in slide support. Do not touch slide or specimen holder.
• Place cartridge upright and support with holder on slide support.
• Remove screw cap with pasteur pipet bulb.
• Place grid in cartridge specimen side up.
• Replace screw cap turning until it stops. Do not over-tighten or force cap.
• Swing aside holder and lower cartridge.
• Remove pipet bulb.
• Holding slide key, insert slide into air lock tube.
• Check position of pin. Remove key.
• Turn large knurled wheel fully clockwise to seal air lock chamber.
• Press red button to release specimen rod and push in rod to stop position.
• Turn rod clockwise in milled-out ring half the distance of ring. Rod will be in horizontal position.
• When red light below meter for high vacuum/beam current goes out, turn rod through remainder of milled-out ring and press into arrest position.

To Insert Grid into Beam

• Press red button on specimen rod and push into intermediate stop.
• Press red button again and gently push rod to insert cartridge into beam.

To Leave Microscope

• Turn condenser 2 control clockwise to spread beam.
• Turn magnification control to low magnification (1250 - 2500).
• Press filament button. Press red lightening button. Remove grid.
• Place holder in microscope in arrest position.

To Turn Off Microscope

• Follow procedure for leaving microscope.
• Press I/O button.

  If power gowe off during operation, press I/O button. Do not remove specimen from microscope.

ALIGNMENT INSTRUCTIONS

• Insert holey grid into arrest position.
• Set magnification at 6300 with magnification lens control.
• Select desired kV.
• Press green lightening button.
• Press filament button.

Filament Alignment

• Turn condenser lens coarse and fine controls to obtain crossover image.
• Move beam with one pair of beam alignment controls to position on screen below post.
• Undersatruate filament by turning filament heating control counter-clockwise until image appears as a center spot with 2 parentheses (CBS eye).
• Move each pair of beam alignment controls (pair of X controls and pair of Y controls) until parentheses around center spot are equal in position, size and intensity.
• Saturate the filament by turning the filament heating control clockwise slowly until crossover image is solid spot. Filament image should not shift position during saturation.

Condenser Aperture Alignment

• Turn condenser 2 lens fine control clockwise until any edge of screen is illuminated.
• Center beam around edge of screen with beam alignment controls.
• Turn condenser 2 lens fine control counter-clockwise through crossover until any edge of screen is illuminated. If edge of screen is not illuminated evenly, move beam half the distance to the dark edge with condenser aperture controls. Repeat steps 1 and 2 until edge of screen is illuminated evenly when beam is overfocused or underfocused from crossover.

Correction of Astigmatism in Condenser 2 Lens

• Turn condenser 2 lens fine control clockwise from crossover until most of large screen is illuminated.
• If beam is not circular, adjust condenser 2 stigmator controls until beam appears circular.

Objective Aperture Alignment

• Insert holey grid into beam path.
• Insert projection 1 aperture to first or second position.
• Center image of aperture around post in screen with projection 1 aperture controls.
• Turn image/diffraction control 1 click to right.
• Focus image of diffraction spot with diffraction pattern focusing control.
• Center image of objective aperture around diffraction spot with objective aperture controls.

Correction of Astigmatism in Projection 1 Lens

• Remove projection 1 aperture from beam.
• Move grid with grid controls until beam is in center of a grid square.
• Defocus image of diffraction spot with diffraction pattern focusing control until star pattern appears.
• Adjust star pattern with projection 1 stigmator controls (P1 stigm wheels) to achieve smallest spot and most uniform pattern.
• Turn image/diffraction control back to image mode.

Correction of Astigmatism in Objective Lens

• Turn condenser lens control counter-clockwise to crossover.
• Center beam around post with beam alignment controls.
• Turn magnification control to 100,000 or 125,000.
• Center image of small, round hole around mark on post.
• Adjust objective lens controls until image is slightly overfocused.
• Image of hole will have black ring (Fresnel fringe) around it.
• Turn objective lens fine control counter-clockwise toward focus while watching the black ring.
• If black ring does not disappear around edge of hole evenly, adjust objective lens fine control until ring disappears on one side. Adjust objective lens stigmator controls until ring is same thickness and disappears evenly. If ring disappears during adjustment, overfocus objective lens with fine control until ring reappears. It will be necessary to move objective lens fine control and the objective stigmator controls alternately during process.
• Turn condenser 2 control clockwise to spread beam.
• Turn magnification control to low magnification (1250 - 2500).

PHOTOGRAPHY

To Take Micrograph

• Set letter dial to desired letter key to permit easy identification of micrographs.
• Select magnification according to outer corner marks on screen.
• Center light with beam alignment knobs if necessary.
• Focus image.
• Adjust condenser 2 control to give desired exposure time on meter. Exposure time should be no less than 1 and no more than 5 sec.
• Press green start button. Exposure cycle is automatic. When green start light reappears, exposure is complete.
• If green start light is not illuminated or fails to reappear after exposure, the camera is empty.

To Load Camera

• Open nitrogen tank by turning knob on top of tank. Do not turn flow control on side of regulator.
• Press Cam button to in position.
• Turn off room light if on.
• Turn off microscope white lights and turn on red lights by pressing red light (circle with X) button on upper right panel.
• Open desiccator.
  • Make sure Desicc key is pressed in and illuminated.
  • Lift ventilation button on top of desiccator to vent.
  • Lift lid of desiccator.
• When magazine lid is loose, turn off nitrogen.
• Pull down frame and remove exposed-film container. Remove film holders and replace container.
• Remove magazine lid and replace empty magazine with full magazine from desiccator. Press down on sides of magazine to release film holders. Replace magazine lid.
• Remove exposed film from film holders and place in light-tight container.
• Press sides of magazine in to lock film supports in place.
• Remove film from package toughing only edges of the film.
• With emulsion side up (light side, notch in upper right corner of film), slide one sheet of film into each holder making sure the film is anchored under rim of holder on all three sides.
• Place loaded film holder in magazine. Make sure film holders are flat and not angled in magazine. There should be 28 film holders in each magazine.
• Check supply of film in desiccator. If you are using last package, replenish supply of film in desiccator. Obtain 1 or 2 film boxes from coldroom across from EM Lab. Remove film packages from box and open by tearing off seal. Remove 2 cardboard supports from packages. Turn foil cover over to close package.
• Check beaker of phosphorus pentoxide in desiccator. If it is wet or liquid, replace it.
  • Place old beaker under hood.
  • Obtain 600-ml beaker from EM drawer in EM Lab.
  • Under hood, place small amount of phosphorus pentoxide in beaker.
• Close desiccator lid. Release Cam button to evacuate camera chamber. Wait until red light goes out (about 3 min).
• Release Desicc button to evacuate desiccator.
• When red light goes out, press in desicc button. This takes 10-15 min, but may take longer if desiccator contains new film.

CARTRIDGES

Gonitometer Stage

• Make sure any cartridge in microscope is in arrest position in air lock.
• Turn on goniometer
• Turn Rem P (remove position ) on. Light must be illuminated before specimen holder is inserted or removed. LED readings should be:
  • Tilt: 90.8
  • Rotate: 1242
• Select cartridge type with cartridge selector.
• Make sure stage controls are set at 500 or in center.
• Remove objective aperture if necessary. Insert cartridge into column.
• Turn on high voltage and filament. Release Rem P Button.
• Press Z ero P (zero position) button. Wait until Tilt and Rotate displays read 0. Release Zero P button.
• Select desired area of specimen.

• To remove any holder
  • Press Rem P button.
  • When LED readings for tilt and rotate return to readings for Rem p, pull holder into arrest position in air lock.
  • Turn off goniometer.
  • Remove holder from microscope.

 

• For goniometer cartridge
  • To place grid in goniometer cartridge
    • Turn tilt drive on manual drive ring to left stop position and rotate drive fully counter-clockwise.
    • Holding cartridge with lens paper or lent-free cloth (or wearing gloves), place cartridge in manual drive ring with screws on cartridge visible in notches on ring and edge aligned with mark.
    • Turn drives until grid holder is slightly tilted and notch aligned with orange rod. Place grid in holder. Hold clip with forceps and press slightly to right to secure clip against grid.
    • Turn tilt drive on manual drive ring to left stop position and rotate drive fully counter-clockwise. Grid holder should be fully tilted and counter-clockwise
    • Remove manual drive ring.
    • Holding cartridge with lens paper or lent-free cloth (or wearing gloves), place cartridge in slide holder.
  • To operate cartridge
    • Select desired magnification.
    • Move object to center.
    • Press left foot pedal to rotate image. If specimen is to be tilted, rotate specimen until object can be center in desired orientation with left stage control. The direction of the tilt axis corresponds to the specimen movement when operating the right stage control.
    • Press right foot pedal to tilt specimen.
  • To determine mganification of image using goniometer cartridge
    • With image focused, record reading for objective lens current on meter.
    • Subtract reading for objective lens current on meter from normal reading for objective lens with standard specimen holder (DI).
    • Locate Mx for DI at set kV on chart. Multiply standard magnification from magnification chart by value for Mx.

     

• For tensile cartridge
  • Glue specimen foil between jaws of cartridge.
  • Set kV to 80 or 100
  • Insert 60-mm objective aperture
  • Move toggle switch on goniometer to V var position.
  • With Rem p depressed, LED display for tilt will read 32.7
  • Turn speed control (right-hand black knob near V switch) fully counter-clockwise. To increase rate of stretching, turn speed control clockwise.
  • View initially at 100X. Set mag to 100X with image/diffraction knob and focus with diffraction pattern focusing knob.
  • To initiate tension, press right side of right foot pedal. Each unit on LED meter for tilt = change of 0.55 mm. Holder will allow maximium stretching of 20% or LED reading of 91.
  • To release tension, press left side of right foot pedal.

 

• For triple cartridge
  • Right foot pedal changes specimen.

MODES OF OPERATION

Darkfield

• Turn beam tilt on.
• Move tilt ( x or y) control until brightfield image just disappears.

Electron Diffraction

• Selected Area Diffraction
  • Withdraw objective aperture.
  • Set magnification to 6300.
  • Insert and center intermediate aperture.
    • 400-mm aperture defined diffracting specimen area of 8 mm
    • 200-mm aperture defined diffracting specimen area of 4mm
    • 100-mm aperture defined diffracting specimen area of 1mm
  • Turn image/diffraction switch to diffraction.
  • Focus diffraction spsot.
  • If necessary, correct for astigmatism with P1 stigmator.
  • Release C1 and C2 buttons to turn off lenses.
  • Select magnification to give desired image size according to intensity and position of reflections.
  • Swing in central beam stop with knob on left side behind viewing chamber to cover zero order diffraction spot.
  • Focus image with diffraction focus control.

 

• Long-angle Diffraction
  • Set magnification ot 6300.
  • Select 30-mm objective aperture and center.
  • Turn image/diffraction swwitch to diffraction.
  • Focus diffraction sspot.
  • Turn condenser 1 lens control to 4 or 5.
  • Release C2 and Obj buttons to turn off lenses
  • Select magnification to give desired image size according to intensity and position of reflections.
  • Swing in central beam stop with knob on left side behind viewing chamber.
  • Focus image with diffraction focus control.