Difference: Jeol1230Instructions (1 vs. 30)

Revision 3026 Feb 2015 - Main.BillRice

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Added:
>
>

Log in

  • User Name: su
  • Password: 1230
 

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Monitor Vacuum condition:
    • Go to "FR Panel" "monitor" "vacuum conditions"
    • When SPC value is <50 insert holder by turning the holder slowly clockwise 1/8 turn to the 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Grasp holder firmly then turn the holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam
    sampleinsertion.jpg

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%
  • Be patient, it takes about one minute for the beam to appear

Condenser Aperture Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right click mouse button and drag the beam to central position
  • Spread Beam (over clockwise) with "Brightness" knob (F)
  • If it does not spread evenly, center with the condenser aperture x-y controls
  • condense beam with "Brightness" (F)
  • If beam is not centered, repeat
  • The beam should expand/ contract concentrically around the beam centre as the brightness is adjusted

Objective Aperture Alignment

  • Select Diffraction mode by pressing "Diffraction" (F)
  • Set the camera length to 150cm
  • Condense beam with "Brightness" (F)
  • Focus beam with "Diff focus" (F)
  • Insert objective aperture
  • Center aperture with x-y controls

Z Height Adjustment

  • On the menu, go to Monitor, then enter Lens condition sub-menu.
  • By adjusting the focus, set the objective lens current to 4.7.
  • Enter Z cont mode, then click AUTO to set specimen eucentric height.

Diffraction

  • Select Diffraction button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Switch the beam is off
  • Switch the toggle to "Air" (G)
  • Monitor Vacuum condition:
    • Go to "FR Panel" "monitor" "vacuum conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out the specimen holder to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessary the SPC is not vented

1230 ShutDown Checklist

FilamentExchange

SerialEM Operation

Startup

  1. Launch Digital Micrograph
  2. Launch Temserver20xx.exe
  3. Launch your copy of SerialEM

Montaging

  • To do a low-mag montage, go to 100X or 80X, fully expand beam, take a test Record image to be sure images are OK
  • Go to Navigator menu, choose to Open
  • Go to Navigator menu, choose Setup Full Montage
  • Choose to save all header info, save file as FILENAME.st
  • Increase overlap percent to 20 (3X3 images for full montage at 100X)
  • Click Start on montage menu
  • Montage will begin. If it appears to get stuck, quickly toggle mags
  • When done, click YES to make a new map
  • Try checking and unchecking Treat as very sloppy montage -- sometimes it is better, sometimes worse
  • Go to File menu, click close to close the file for writing (do not want to add to this file)

Alignment of Montages

  • First map is registration 1
  • Make a new montage, either at a different mag or at a different grid orientation
  • Follow instructions above, but before you click start, CHANGE CURRENT REGISTRATION TO 2
  • Make the new map -- you now have old map in registration 1 and new in registration 2
  • Need at least 3 points to align the maps (determine a shift/rotation matrix)
  • Choose three obvious points, click Add Points, and add points. When done, click stop
  • Mark each point as a Registration point, numbered 1, 2 and 3
  • Re-open first map by double clicking
  • CHANGE CURRENT REGISTRATION BACK TO 1
  • Click Add Points, and add the three corresponding points on the old map
  • Mark these as Registration Points , again 1,2,3, IN THE SAME ORDER AS ON MAP 2
  • Change current registration back to 2, and re-load new map
  • Under Navigator menu, choose Transform items
  • Map 1 is now transformed onto map 2

Acquiring many areas using polygon montage

  • Change to desired new magnification (say 4000X )
  • Take Record image to test beam and image settings
  • In low mag map, choose Add Polygon and outline a grid square
  • With Polygon highlighted, go to Navigator menu and choose Setup Polygon Montage
  • Increase overlap to 15%-20%, and also choose to Move Stage
  • Change current registration as described above, and click start
  • When polygon montage is done, create a new map and align it to old map as described above
  • DO NOT CLOSE THE NEW FILE
  • In low mag map, decide which squares are to be acquired, and add a point in the middle of each
  • Mark each new point as ACQUIRE
  • Under Navigator menu, choose Acquire Areas
  • A polygon montage, centered at each point, will now be acquired.

Running a script at many points

  • Under Macro mwnu, edit your script as desired
  • In a map, choose to add points, and add them to desired areas
  • Mark each point as ACQUIRE
  • Under Navigator menu, choose Run Macro at areas
* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

META FILEATTACHMENT attachment="sampleinsertion.jpg" attr="" comment="" date="1165000068" name="sampleinsertion.jpg" path="sampleinsertion.jpg" size="194485" stream="sampleinsertion.jpg" user="Main.KdDerr" version="1"

Revision 2920 Feb 2008 - Main.BillRice

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Monitor Vacuum condition:
    • Go to "FR Panel" "monitor" "vacuum conditions"
    • When SPC value is <50 insert holder by turning the holder slowly clockwise 1/8 turn to the 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Grasp holder firmly then turn the holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam
    sampleinsertion.jpg

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%
  • Be patient, it takes about one minute for the beam to appear

Condenser Aperture Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right click mouse button and drag the beam to central position
  • Spread Beam (over clockwise) with "Brightness" knob (F)
  • If it does not spread evenly, center with the condenser aperture x-y controls
  • condense beam with "Brightness" (F)
  • If beam is not centered, repeat
  • The beam should expand/ contract concentrically around the beam centre as the brightness is adjusted

Objective Aperture Alignment

  • Select Diffraction mode by pressing "Diffraction" (F)
  • Set the camera length to 150cm
  • Condense beam with "Brightness" (F)
  • Focus beam with "Diff focus" (F)
  • Insert objective aperture
  • Center aperture with x-y controls

Z Height Adjustment

  • On the menu, go to Monitor, then enter Lens condition sub-menu.
  • By adjusting the focus, set the objective lens current to 4.7.
  • Enter Z cont mode, then click AUTO to set specimen eucentric height.

Diffraction

  • Select Diffraction button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Switch the beam is off
  • Switch the toggle to "Air" (G)
  • Monitor Vacuum condition:
    • Go to "FR Panel" "monitor" "vacuum conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out the specimen holder to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessary the SPC is not vented

1230 ShutDown Checklist

FilamentExchange

SerialEM Operation

Startup

  1. Launch Digital Micrograph
  2. Launch Temserver20xx.exe
  3. Launch your copy of SerialEM

Montaging

  • To do a low-mag montage, go to 100X or 80X, fully expand beam, take a test Record image to be sure images are OK
  • Go to Navigator menu, choose to Open
  • Go to Navigator menu, choose Setup Full Montage
  • Choose to save all header info, save file as FILENAME.st
  • Increase overlap percent to 20 (3X3 images for full montage at 100X)
  • Click Start on montage menu
  • Montage will begin. If it appears to get stuck, quickly toggle mags
  • When done, click YES to make a new map
  • Try checking and unchecking Treat as very sloppy montage -- sometimes it is better, sometimes worse
  • Go to File menu, click close to close the file for writing (do not want to add to this file)

Alignment of Montages

  • First map is registration 1
  • Make a new montage, either at a different mag or at a different grid orientation
  • Follow instructions above, but before you click start, CHANGE CURRENT REGISTRATION TO 2
  • Make the new map -- you now have old map in registration 1 and new in registration 2
  • Need at least 3 points to align the maps (determine a shift/rotation matrix)
  • Choose three obvious points, click Add Points, and add points. When done, click stop
  • Mark each point as a Registration point, numbered 1, 2 and 3
  • Re-open first map by double clicking
  • CHANGE CURRENT REGISTRATION BACK TO 1
  • Click Add Points, and add the three corresponding points on the old map
  • Mark these as Registration Points , again 1,2,3, IN THE SAME ORDER AS ON MAP 2
  • Change current registration back to 2, and re-load new map
  • Under Navigator menu, choose Transform items
  • Map 1 is now transformed onto map 2
Added:
>
>

Acquiring many areas using polygon montage

  • Change to desired new magnification (say 4000X )
  • Take Record image to test beam and image settings
  • In low mag map, choose Add Polygon and outline a grid square
  • With Polygon highlighted, go to Navigator menu and choose Setup Polygon Montage
  • Increase overlap to 15%-20%, and also choose to Move Stage
  • Change current registration as described above, and click start
  • When polygon montage is done, create a new map and align it to old map as described above
  • DO NOT CLOSE THE NEW FILE
  • In low mag map, decide which squares are to be acquired, and add a point in the middle of each
  • Mark each new point as ACQUIRE
  • Under Navigator menu, choose Acquire Areas
  • A polygon montage, centered at each point, will now be acquired.

Running a script at many points

  • Under Macro mwnu, edit your script as desired
  • In a map, choose to add points, and add them to desired areas
  • Mark each point as ACQUIRE
  • Under Navigator menu, choose Run Macro at areas
 * Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

META FILEATTACHMENT attachment="sampleinsertion.jpg" attr="" comment="" date="1165000068" name="sampleinsertion.jpg" path="sampleinsertion.jpg" size="194485" stream="sampleinsertion.jpg" user="Main.KdDerr" version="1"

Revision 2820 Feb 2008 - Main.BillRice

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Monitor Vacuum condition:
    • Go to "FR Panel" "monitor" "vacuum conditions"
    • When SPC value is <50 insert holder by turning the holder slowly clockwise 1/8 turn to the 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Grasp holder firmly then turn the holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam
    sampleinsertion.jpg

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%
  • Be patient, it takes about one minute for the beam to appear

Condenser Aperture Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right click mouse button and drag the beam to central position
  • Spread Beam (over clockwise) with "Brightness" knob (F)
  • If it does not spread evenly, center with the condenser aperture x-y controls
  • condense beam with "Brightness" (F)
  • If beam is not centered, repeat
  • The beam should expand/ contract concentrically around the beam centre as the brightness is adjusted

Objective Aperture Alignment

  • Select Diffraction mode by pressing "Diffraction" (F)
  • Set the camera length to 150cm
  • Condense beam with "Brightness" (F)
  • Focus beam with "Diff focus" (F)
  • Insert objective aperture
  • Center aperture with x-y controls

Z Height Adjustment

  • On the menu, go to Monitor, then enter Lens condition sub-menu.
  • By adjusting the focus, set the objective lens current to 4.7.
  • Enter Z cont mode, then click AUTO to set specimen eucentric height.

Diffraction

  • Select Diffraction button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Switch the beam is off
  • Switch the toggle to "Air" (G)
  • Monitor Vacuum condition:
    • Go to "FR Panel" "monitor" "vacuum conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out the specimen holder to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessary the SPC is not vented

1230 ShutDown Checklist

FilamentExchange

Deleted:
<
<
 
Added:
>
>

SerialEM Operation

Startup

  1. Launch Digital Micrograph
  2. Launch Temserver20xx.exe
  3. Launch your copy of SerialEM

Montaging

  • To do a low-mag montage, go to 100X or 80X, fully expand beam, take a test Record image to be sure images are OK
  • Go to Navigator menu, choose to Open
  • Go to Navigator menu, choose Setup Full Montage
  • Choose to save all header info, save file as FILENAME.st
  • Increase overlap percent to 20 (3X3 images for full montage at 100X)
  • Click Start on montage menu
  • Montage will begin. If it appears to get stuck, quickly toggle mags
  • When done, click YES to make a new map
  • Try checking and unchecking Treat as very sloppy montage -- sometimes it is better, sometimes worse
  • Go to File menu, click close to close the file for writing (do not want to add to this file)

Alignment of Montages

  • First map is registration 1
  • Make a new montage, either at a different mag or at a different grid orientation
  • Follow instructions above, but before you click start, CHANGE CURRENT REGISTRATION TO 2
  • Make the new map -- you now have old map in registration 1 and new in registration 2
  • Need at least 3 points to align the maps (determine a shift/rotation matrix)
  • Choose three obvious points, click Add Points, and add points. When done, click stop
  • Mark each point as a Registration point, numbered 1, 2 and 3
  • Re-open first map by double clicking
  • CHANGE CURRENT REGISTRATION BACK TO 1
  • Click Add Points, and add the three corresponding points on the old map
  • Mark these as Registration Points , again 1,2,3, IN THE SAME ORDER AS ON MAP 2
  • Change current registration back to 2, and re-load new map
  • Under Navigator menu, choose Transform items
  • Map 1 is now transformed onto map 2
 * Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

META FILEATTACHMENT attachment="sampleinsertion.jpg" attr="" comment="" date="1165000068" name="sampleinsertion.jpg" path="sampleinsertion.jpg" size="194485" stream="sampleinsertion.jpg" user="Main.KdDerr" version="1"

Revision 2727 Jul 2007 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Monitor Vacuum condition:
    • Go to "FR Panel" "monitor" "vacuum conditions"
    • When SPC value is <50 insert holder by turning the holder slowly clockwise 1/8 turn to the 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Grasp holder firmly then turn the holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam
    sampleinsertion.jpg

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%
  • Be patient, it takes about one minute for the beam to appear

Condenser Aperture Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right click mouse button and drag the beam to central position
  • Spread Beam (over clockwise) with "Brightness" knob (F)
  • If it does not spread evenly, center with the condenser aperture x-y controls
  • condense beam with "Brightness" (F)
  • If beam is not centered, repeat
  • The beam should expand/ contract concentrically around the beam centre as the brightness is adjusted

Objective Aperture Alignment

  • Select Diffraction mode by pressing "Diffraction" (F)
  • Set the camera length to 150cm
  • Condense beam with "Brightness" (F)
  • Focus beam with "Diff focus" (F)
  • Insert objective aperture
  • Center aperture with x-y controls

Z Height Adjustment

  • On the menu, go to Monitor, then enter Lens condition sub-menu.
  • By adjusting the focus, set the objective lens current to 4.7.
  • Enter Z cont mode, then click AUTO to set specimen eucentric height.

Diffraction

  • Select Diffraction button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Switch the beam is off
  • Switch the toggle to "Air" (G)
  • Monitor Vacuum condition:
    • Go to "FR Panel" "monitor" "vacuum conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out the specimen holder to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessary the SPC is not vented

1230 ShutDown Checklist

Added:
>
>

FilamentExchange

 

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

META FILEATTACHMENT attachment="sampleinsertion.jpg" attr="" comment="" date="1165000068" name="sampleinsertion.jpg" path="sampleinsertion.jpg" size="194485" stream="sampleinsertion.jpg" user="Main.KdDerr" version="1"

Revision 2607 Mar 2007 - Main.AlistairSiebert

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Monitor Vacuum condition:
    • Go to "FR Panel" "monitor" "vacuum conditions"
    • When SPC value is <50 insert holder by turning the holder slowly clockwise 1/8 turn to the 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Grasp holder firmly then turn the holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam
    sampleinsertion.jpg

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%
  • Be patient, it takes about one minute for the beam to appear

Condenser Aperture Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right click mouse button and drag the beam to central position
  • Spread Beam (over clockwise) with "Brightness" knob (F)
Changed:
<
<
  • If it does not spread evenly, center with the condenser aperture x-y
>
>
  • If it does not spread evenly, center with the condenser aperture x-y controls
 
  • condense beam with "Brightness" (F)
  • If beam is not centered, repeat
  • The beam should expand/ contract concentrically around the beam centre as the brightness is adjusted

Objective Aperture Alignment

Changed:
<
<
  • Select Diffraction mode (F)
>
>
  • Select Diffraction mode by pressing "Diffraction" (F)
 
  • Set the camera length to 150cm
Changed:
<
<
  • Condense beam with Brightness (F)
  • Focus beam with Diffraction focus (F)
>
>
  • Condense beam with "Brightness" (F)
  • Focus beam with "Diff focus" (F)
 
  • Insert objective aperture
Changed:
<
<
  • Center beam within aperture with diffraction aperture x-y align
>
>
  • Center aperture with x-y controls
 

Z Height Adjustment

Changed:
<
<
  • Set the objective lens current to 4.7 (sub-menu?)
  • Go into x mode and
  • (unfinished)
>
>
  • On the menu, go to Monitor, then enter Lens condition sub-menu.
  • By adjusting the focus, set the objective lens current to 4.7.
  • Enter Z cont mode, then click AUTO to set specimen eucentric height.
 

Diffraction

  • Select Diffraction button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Switch the beam is off
  • Switch the toggle to "Air" (G)
  • Monitor Vacuum condition:
    • Go to "FR Panel" "monitor" "vacuum conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out the specimen holder to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessary the SPC is not vented

1230 ShutDown Checklist

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

META FILEATTACHMENT attachment="sampleinsertion.jpg" attr="" comment="" date="1165000068" name="sampleinsertion.jpg" path="sampleinsertion.jpg" size="194485" stream="sampleinsertion.jpg" user="Main.KdDerr" version="1"

Revision 2507 Mar 2007 - Main.AlistairSiebert

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Monitor Vacuum condition:
Changed:
<
<
    • Go to "FR Panel" "monitor" "vacuum Conditions"
    • SPC value needs to be 50 or lower before insertion
>
>
    • Go to "FR Panel" "monitor" "vacuum conditions"
    • When SPC value is <50 insert holder by turning the holder slowly clockwise 1/8 turn to the 1st stop
Deleted:
<
<
    • If sample rod was left out overnight it may time out
  • Turn holder slowly clockwise 1/8 turn to 1st stop
 
  • Slowly allow the vacuum to pull the holder in
Changed:
<
<
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
>
>
  • Grasp holder firmly then turn the holder slowly clockwise 1/4 turn to 2nd stop
 
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam
    sampleinsertion.jpg

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%
  • Be patient, it takes about one minute for the beam to appear
Changed:
<
<

Condenser Lens Alignment

>
>

Condenser Aperture Alignment

 
  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
Changed:
<
<
  • Right Click mouse button and drag to center beam
>
>
  • Right click mouse button and drag the beam to central position
 
  • Spread Beam (over clockwise) with "Brightness" knob (F)
Changed:
<
<
  • If it does not spread evenly center with Condenser aperture x-y
>
>
  • If it does not spread evenly, center with the condenser aperture x-y
 
  • condense beam with "Brightness" (F)
Changed:
<
<
    • If beam is not centered, repeat
>
>
  • If beam is not centered, repeat
Added:
>
>
  • The beam should expand/ contract concentrically around the beam centre as the brightness is adjusted
 
Changed:
<
<

Alignment of Objective Aperture

  • Select Diffraction button (F)
>
>

Objective Aperture Alignment

  • Select Diffraction mode (F)
 
  • Set the camera length to 150cm
  • Condense beam with Brightness (F)
Changed:
<
<
  • Focus beam with Dif focus (F)
>
>
  • Focus beam with Diffraction focus (F)
 
  • Insert objective aperture
  • Center beam within aperture with diffraction aperture x-y align

Z Height Adjustment

  • Set the objective lens current to 4.7 (sub-menu?)
Changed:
<
<
  • Go into x mode and (unfinished)
>
>
  • Go into x mode and
Added:
>
>
  • (unfinished)
 

Diffraction

  • Select Diffraction button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Switch the beam is off
  • Switch the toggle to "Air" (G)
  • Monitor Vacuum condition:
    • Go to "FR Panel" "monitor" "vacuum conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out the specimen holder to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessary the SPC is not vented

1230 ShutDown Checklist

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

META FILEATTACHMENT attachment="sampleinsertion.jpg" attr="" comment="" date="1165000068" name="sampleinsertion.jpg" path="sampleinsertion.jpg" size="194485" stream="sampleinsertion.jpg" user="Main.KdDerr" version="1"

Revision 2407 Mar 2007 - Main.AlistairSiebert

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
Changed:
<
<
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
>
>
  • Monitor Vacuum condition:
    • Go to "FR Panel" "monitor" "vacuum Conditions"
 
    • SPC value needs to be 50 or lower before insertion
Changed:
<
<
    • If sample rod was out in atmosphere overnight it may time out
>
>
    • If sample rod was left out overnight it may time out
 
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam
    sampleinsertion.jpg

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%
Changed:
<
<
  • Be patient it takes about one minute for the beam to appear
>
>
  • Be patient, it takes about one minute for the beam to appear
 

Condenser Lens Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
Changed:
<
<
  • Right Click mouse and drag to center beam
  • Spread Beam (over clockwise) with "Brightness" Knob (F)
>
>
  • Right Click mouse button and drag to center beam
  • Spread Beam (over clockwise) with "Brightness" knob (F)
 
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat
Changed:
<
<

Alignment of DIffration Aperture

>
>

Alignment of Objective Aperture

 
  • Select Diffraction button (F)
Added:
>
>
  • Set the camera length to 150cm
 
  • Condense beam with Brightness (F)
Changed:
<
<
  • Sharpen beam with Dif Focus (F)
  • Insert Diffraction aperture
>
>
  • Focus beam with Dif focus (F)
  • Insert objective aperture
 
  • Center beam within aperture with diffraction aperture x-y align

Z Height Adjustment

Added:
>
>
  • Set the objective lens current to 4.7 (sub-menu?)
  • Go into x mode and (unfinished)
 

Diffraction

Changed:
<
<
  • Select Diffration button (F)
>
>
  • Select Diffraction button (F)
 
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

Changed:
<
<
  • Make sure the beam is off
>
>
  • Switch the beam is off
 
  • Switch the toggle to "Air" (G)
Changed:
<
<
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
>
>
  • Monitor Vacuum condition:
    • Go to "FR Panel" "monitor" "vacuum conditions"
 
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
Changed:
<
<
  • Pull out agian to the 2nd stop
>
>
  • Pull out the specimen holder to the 2nd stop
 
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
Changed:
<
<
    • If the use of force is necessiary the SPC is not vented
>
>
    • If the use of force is necessary the SPC is not vented
 

1230 ShutDown Checklist

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

META FILEATTACHMENT attachment="sampleinsertion.jpg" attr="" comment="" date="1165000068" name="sampleinsertion.jpg" path="sampleinsertion.jpg" size="194485" stream="sampleinsertion.jpg" user="Main.KdDerr" version="1"

Revision 2306 Dec 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • SPC value needs to be 50 or lower before insertion
    • If sample rod was out in atmosphere overnight it may time out
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam
    sampleinsertion.jpg

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%
  • Be patient it takes about one minute for the beam to appear

Condenser Lens Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right Click mouse and drag to center beam
  • Spread Beam (over clockwise) with "Brightness" Knob (F)
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat

Alignment of DIffration Aperture

  • Select Diffraction button (F)
  • Condense beam with Brightness (F)
  • Sharpen beam with Dif Focus (F)
  • Insert Diffraction aperture
  • Center beam within aperture with diffraction aperture x-y align

Z Height Adjustment

Diffraction

  • Select Diffration button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Make sure the beam is off
  • Switch the toggle to "Air" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessiary the SPC is not vented
Changed:
<
<

1230ShutDown Checklist

>
>

1230 ShutDown Checklist

 

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

META FILEATTACHMENT attachment="sampleinsertion.jpg" attr="" comment="" date="1165000068" name="sampleinsertion.jpg" path="sampleinsertion.jpg" size="194485" stream="sampleinsertion.jpg" user="Main.KdDerr" version="1"

Revision 2206 Dec 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • SPC value needs to be 50 or lower before insertion
    • If sample rod was out in atmosphere overnight it may time out
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam
    sampleinsertion.jpg

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%
  • Be patient it takes about one minute for the beam to appear

Condenser Lens Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right Click mouse and drag to center beam
  • Spread Beam (over clockwise) with "Brightness" Knob (F)
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat

Alignment of DIffration Aperture

  • Select Diffraction button (F)
  • Condense beam with Brightness (F)
  • Sharpen beam with Dif Focus (F)
  • Insert Diffraction aperture
  • Center beam within aperture with diffraction aperture x-y align

Z Height Adjustment

Diffraction

  • Select Diffration button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Make sure the beam is off
  • Switch the toggle to "Air" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessiary the SPC is not vented
Changed:
<
<

ShutDown Checklist

>
>

1230ShutDown Checklist

 

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

META FILEATTACHMENT attachment="sampleinsertion.jpg" attr="" comment="" date="1165000068" name="sampleinsertion.jpg" path="sampleinsertion.jpg" size="194485" stream="sampleinsertion.jpg" user="Main.KdDerr" version="1"

Revision 2106 Dec 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • SPC value needs to be 50 or lower before insertion
    • If sample rod was out in atmosphere overnight it may time out
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam
    sampleinsertion.jpg

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%
  • Be patient it takes about one minute for the beam to appear

Condenser Lens Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right Click mouse and drag to center beam
  • Spread Beam (over clockwise) with "Brightness" Knob (F)
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat

Alignment of DIffration Aperture

  • Select Diffraction button (F)
  • Condense beam with Brightness (F)
  • Sharpen beam with Dif Focus (F)
  • Insert Diffraction aperture
  • Center beam within aperture with diffraction aperture x-y align

Z Height Adjustment

Diffraction

  • Select Diffration button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Make sure the beam is off
  • Switch the toggle to "Air" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessiary the SPC is not vented
Added:
>
>

ShutDown Checklist

 

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

META FILEATTACHMENT attachment="sampleinsertion.jpg" attr="" comment="" date="1165000068" name="sampleinsertion.jpg" path="sampleinsertion.jpg" size="194485" stream="sampleinsertion.jpg" user="Main.KdDerr" version="1"

Revision 2001 Dec 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • SPC value needs to be 50 or lower before insertion
    • If sample rod was out in atmosphere overnight it may time out
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam
    sampleinsertion.jpg

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%
  • Be patient it takes about one minute for the beam to appear

Condenser Lens Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right Click mouse and drag to center beam
  • Spread Beam (over clockwise) with "Brightness" Knob (F)
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat

Alignment of DIffration Aperture

  • Select Diffraction button (F)
  • Condense beam with Brightness (F)
  • Sharpen beam with Dif Focus (F)
  • Insert Diffraction aperture
  • Center beam within aperture with diffraction aperture x-y align

Z Height Adjustment

Diffraction

  • Select Diffration button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Make sure the beam is off
  • Switch the toggle to "Air" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessiary the SPC is not vented

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

Deleted:
<
<		 * sampleinsertion.jpg:
sampleinsertion.jpg
 
META FILEATTACHMENT attachment="sampleinsertion.jpg" attr="" comment="" date="1165000068" name="sampleinsertion.jpg" path="sampleinsertion.jpg" size="194485" stream="sampleinsertion.jpg" user="Main.KdDerr" version="1"

Revision 1901 Dec 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • SPC value needs to be 50 or lower before insertion
    • If sample rod was out in atmosphere overnight it may time out
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam
Changed:
<
<
  • sampleinsertion.jpg:
>
>
  sampleinsertion.jpg

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%
  • Be patient it takes about one minute for the beam to appear

Condenser Lens Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right Click mouse and drag to center beam
  • Spread Beam (over clockwise) with "Brightness" Knob (F)
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat

Alignment of DIffration Aperture

  • Select Diffraction button (F)
  • Condense beam with Brightness (F)
  • Sharpen beam with Dif Focus (F)
  • Insert Diffraction aperture
  • Center beam within aperture with diffraction aperture x-y align

Z Height Adjustment

Diffraction

  • Select Diffration button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Make sure the beam is off
  • Switch the toggle to "Air" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessiary the SPC is not vented

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006 * sampleinsertion.jpg:
sampleinsertion.jpg

META FILEATTACHMENT attachment="sampleinsertion.jpg" attr="" comment="" date="1165000068" name="sampleinsertion.jpg" path="sampleinsertion.jpg" size="194485" stream="sampleinsertion.jpg" user="Main.KdDerr" version="1"

Revision 1801 Dec 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • SPC value needs to be 50 or lower before insertion
    • If sample rod was out in atmosphere overnight it may time out
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam
Changed:
<
<		 * sampleinsertion.jpg:
>
>
  • sampleinsertion.jpg:
  sampleinsertion.jpg

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%
  • Be patient it takes about one minute for the beam to appear

Condenser Lens Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right Click mouse and drag to center beam
  • Spread Beam (over clockwise) with "Brightness" Knob (F)
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat

Alignment of DIffration Aperture

  • Select Diffraction button (F)
  • Condense beam with Brightness (F)
  • Sharpen beam with Dif Focus (F)
  • Insert Diffraction aperture
  • Center beam within aperture with diffraction aperture x-y align

Z Height Adjustment

Diffraction

  • Select Diffration button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Make sure the beam is off
  • Switch the toggle to "Air" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessiary the SPC is not vented

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006 * sampleinsertion.jpg:
sampleinsertion.jpg

META FILEATTACHMENT attachment="sampleinsertion.jpg" attr="" comment="" date="1165000068" name="sampleinsertion.jpg" path="sampleinsertion.jpg" size="194485" stream="sampleinsertion.jpg" user="Main.KdDerr" version="1"

Revision 1701 Dec 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • SPC value needs to be 50 or lower before insertion
    • If sample rod was out in atmosphere overnight it may time out
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam
Changed:
<
<
>
>		 * sampleinsertion.jpg:
Added:
>
>		 sampleinsertion.jpg
 

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%
  • Be patient it takes about one minute for the beam to appear

Condenser Lens Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right Click mouse and drag to center beam
  • Spread Beam (over clockwise) with "Brightness" Knob (F)
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat

Alignment of DIffration Aperture

  • Select Diffraction button (F)
  • Condense beam with Brightness (F)
  • Sharpen beam with Dif Focus (F)
  • Insert Diffraction aperture
  • Center beam within aperture with diffraction aperture x-y align

Z Height Adjustment

Diffraction

  • Select Diffration button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Make sure the beam is off
  • Switch the toggle to "Air" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessiary the SPC is not vented

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006 * sampleinsertion.jpg:

Changed:
<
<		 sampleinsertion.jpg
>
>		 sampleinsertion.jpg
 
META FILEATTACHMENT attachment="sampleinsertion.jpg" attr="" comment="" date="1165000068" name="sampleinsertion.jpg" path="sampleinsertion.jpg" size="194485" stream="sampleinsertion.jpg" user="Main.KdDerr" version="1"

Revision 1601 Dec 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • SPC value needs to be 50 or lower before insertion
    • If sample rod was out in atmosphere overnight it may time out
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%
  • Be patient it takes about one minute for the beam to appear

Condenser Lens Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right Click mouse and drag to center beam
  • Spread Beam (over clockwise) with "Brightness" Knob (F)
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat

Alignment of DIffration Aperture

  • Select Diffraction button (F)
  • Condense beam with Brightness (F)
  • Sharpen beam with Dif Focus (F)
  • Insert Diffraction aperture
  • Center beam within aperture with diffraction aperture x-y align

Z Height Adjustment

Diffraction

  • Select Diffration button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Make sure the beam is off
  • Switch the toggle to "Air" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessiary the SPC is not vented

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

Added:
>
>		 * sampleinsertion.jpg:
sampleinsertion.jpg

META FILEATTACHMENT attachment="sampleinsertion.jpg" attr="" comment="" date="1165000068" name="sampleinsertion.jpg" path="sampleinsertion.jpg" size="194485" stream="sampleinsertion.jpg" user="Main.KdDerr" version="1"
 

Revision 1501 Dec 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
Changed:
<
<
    • SPC value needs to be 40 or lower and read "Vac. Ready" before insertion
    • If sample rod was out in atmosphere overnight it may take several pumping cycles before it is ready to insert
>
>
    • SPC value needs to be 50 or lower before insertion
    • If sample rod was out in atmosphere overnight it may time out
Deleted:
<
<
  • Watch for green light to go on along with the orange
 
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
Deleted:
<
<
  • Both lights will go out
 
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%
  • Be patient it takes about one minute for the beam to appear

Condenser Lens Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right Click mouse and drag to center beam
  • Spread Beam (over clockwise) with "Brightness" Knob (F)
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat

Alignment of DIffration Aperture

  • Select Diffraction button (F)
  • Condense beam with Brightness (F)
  • Sharpen beam with Dif Focus (F)
  • Insert Diffraction aperture
  • Center beam within aperture with diffraction aperture x-y align

Z Height Adjustment

Diffraction

  • Select Diffration button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Make sure the beam is off
  • Switch the toggle to "Air" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessiary the SPC is not vented

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

Revision 1412 Sep 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • SPC value needs to be 40 or lower and read "Vac. Ready" before insertion
    • If sample rod was out in atmosphere overnight it may take several pumping cycles before it is ready to insert
  • Watch for green light to go on along with the orange
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Both lights will go out
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%
  • Be patient it takes about one minute for the beam to appear

Condenser Lens Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right Click mouse and drag to center beam
  • Spread Beam (over clockwise) with "Brightness" Knob (F)
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat

Alignment of DIffration Aperture

  • Select Diffraction button (F)
  • Condense beam with Brightness (F)
  • Sharpen beam with Dif Focus (F)
  • Insert Diffraction aperture
  • Center beam within aperture with diffraction aperture x-y align
Added:
>
>

Z Height Adjustment

 

Diffraction

  • Select Diffration button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Make sure the beam is off
  • Switch the toggle to "Air" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessiary the SPC is not vented

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

Revision 1312 Sep 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • SPC value needs to be 40 or lower and read "Vac. Ready" before insertion
    • If sample rod was out in atmosphere overnight it may take several pumping cycles before it is ready to insert
  • Watch for green light to go on along with the orange
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Both lights will go out
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%
Added:
>
>
  • Be patient it takes about one minute for the beam to appear
 

Condenser Lens Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right Click mouse and drag to center beam
  • Spread Beam (over clockwise) with "Brightness" Knob (F)
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat

Alignment of DIffration Aperture

  • Select Diffraction button (F)
  • Condense beam with Brightness (F)
  • Sharpen beam with Dif Focus (F)
  • Insert Diffraction aperture
  • Center beam within aperture with diffraction aperture x-y align

Diffraction

  • Select Diffration button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Make sure the beam is off
  • Switch the toggle to "Air" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessiary the SPC is not vented

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

Revision 1224 Aug 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • SPC value needs to be 40 or lower and read "Vac. Ready" before insertion
    • If sample rod was out in atmosphere overnight it may take several pumping cycles before it is ready to insert
  • Watch for green light to go on along with the orange
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Both lights will go out
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%

Condenser Lens Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right Click mouse and drag to center beam
  • Spread Beam (over clockwise) with "Brightness" Knob (F)
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat

Alignment of DIffration Aperture

  • Select Diffraction button (F)
  • Condense beam with Brightness (F)
Added:
>
>
  • Sharpen beam with Dif Focus (F)
 
  • Insert Diffraction aperture
  • Center beam within aperture with diffraction aperture x-y align

Diffraction

  • Select Diffration button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Make sure the beam is off
  • Switch the toggle to "Air" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessiary the SPC is not vented

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

Revision 1124 Aug 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Flip toggle switch to "pump" (G)
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • SPC value needs to be 40 or lower and read "Vac. Ready" before insertion
    • If sample rod was out in atmosphere overnight it may take several pumping cycles before it is ready to insert
  • Watch for green light to go on along with the orange
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Both lights will go out
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%

Condenser Lens Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
Changed:
<
<
  • Right Click mouse to center beam
  • Spread Beam over clockwise with "Brightness" Knob (F)
>
>
  • Right Click mouse and drag to center beam
  • Spread Beam (over clockwise) with "Brightness" Knob (F)
 
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat

Alignment of DIffration Aperture

  • Select Diffraction button (F)
  • Condense beam with Brightness (F)
  • Insert Diffraction aperture
  • Center beam within aperture with diffraction aperture x-y align

Diffraction

  • Select Diffration button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Make sure the beam is off
  • Switch the toggle to "Air" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessiary the SPC is not vented

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

Revision 1024 Aug 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

Added:
>
>
  • Flip toggle switch to "pump" (G)
 
  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
Deleted:
<
<
  • Flip toggle switch to "pump" (G)
 
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • SPC value needs to be 40 or lower and read "Vac. Ready" before insertion
    • If sample rod was out in atmosphere overnight it may take several pumping cycles before it is ready to insert
  • Watch for green light to go on along with the orange
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Both lights will go out
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%

Condenser Lens Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right Click mouse to center beam
  • Spread Beam over clockwise with "Brightness" Knob (F)
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat

Alignment of DIffration Aperture

  • Select Diffraction button (F)
  • Condense beam with Brightness (F)
  • Insert Diffraction aperture
  • Center beam within aperture with diffraction aperture x-y align

Diffraction

  • Select Diffration button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Make sure the beam is off
  • Switch the toggle to "Air" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessiary the SPC is not vented

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

Revision 924 Aug 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Flip toggle switch to "pump" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • SPC value needs to be 40 or lower and read "Vac. Ready" before insertion
    • If sample rod was out in atmosphere overnight it may take several pumping cycles before it is ready to insert
  • Watch for green light to go on along with the orange
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Both lights will go out
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%

Condenser Lens Alignment

  • At 5000x
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right Click mouse to center beam
Changed:
<
<
  • Spread Beam with "Brightness Knob (F)
>
>
  • Spread Beam over clockwise with "Brightness" Knob (F)
 
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat

Alignment of DIffration Aperture

  • Select Diffraction button (F)
  • Condense beam with Brightness (F)
  • Insert Diffraction aperture
  • Center beam within aperture with diffraction aperture x-y align

Diffraction

  • Select Diffration button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Make sure the beam is off
  • Switch the toggle to "Air" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessiary the SPC is not vented

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

Revision 824 Aug 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Flip toggle switch to "pump" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • SPC value needs to be 40 or lower and read "Vac. Ready" before insertion
    • If sample rod was out in atmosphere overnight it may take several pumping cycles before it is ready to insert
  • Watch for green light to go on along with the orange
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Both lights will go out
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%

Condenser Lens Alignment

Added:
>
>
  • At 5000x
 
  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right Click mouse to center beam
  • Spread Beam with "Brightness Knob (F)
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat

Alignment of DIffration Aperture

  • Select Diffraction button (F)
  • Condense beam with Brightness (F)
  • Insert Diffraction aperture
  • Center beam within aperture with diffraction aperture x-y align

Diffraction

  • Select Diffration button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Make sure the beam is off
  • Switch the toggle to "Air" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessiary the SPC is not vented

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

Revision 713 Jun 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Flip toggle switch to "pump" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • SPC value needs to be 40 or lower and read "Vac. Ready" before insertion
    • If sample rod was out in atmosphere overnight it may take several pumping cycles before it is ready to insert
  • Watch for green light to go on along with the orange
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Both lights will go out
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%

Condenser Lens Alignment

  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right Click mouse to center beam
  • Spread Beam with "Brightness Knob (F)
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat

Alignment of DIffration Aperture

  • Select Diffraction button (F)
  • Condense beam with Brightness (F)
  • Insert Diffraction aperture
  • Center beam within aperture with diffraction aperture x-y align

Diffraction

  • Select Diffration button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Make sure the beam is off
  • Switch the toggle to "Air" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessiary the SPC is not vented

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

Revision 613 Jun 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Line up pin on holder with goniometer(G)
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Flip toggle switch to "pump" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • SPC value needs to be 40 or lower and read "Vac. Ready" before insertion
    • If sample rod was out in atmosphere overnight it may take several pumping cycles before it is ready to insert
  • Watch for green light to go on along with the orange
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Both lights will go out
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%

Condenser Lens Alignment

  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right Click mouse to center beam
  • Spread Beam with "Brightness Knob (F)
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat

Alignment of DIffration Aperture

  • Select Diffraction button (F)
Changed:
<
<
  • COndense beam with Brightness (F)
>
>
  • Condense beam with Brightness (F)
 
  • Insert Diffraction aperture
  • Center beam within aperture with diffraction aperture x-y align

Diffraction

  • Select Diffration button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"

Specimen Removal

  • Make sure the beam is off
  • Switch the toggle to "Air" (G)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod (G)
    • If the use of force is necessiary the SPC is not vented

* Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

Revision 513 Jun 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

Changed:
<
<
  • Line up pin on holder with the notched opening
>
>
  • Line up pin on holder with goniometer(G)
 
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
Changed:
<
<
  • Flip toggle switch to "pump" (located below insertion point)
>
>
  • Flip toggle switch to "pump" (G)
 
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • SPC value needs to be 40 or lower and read "Vac. Ready" before insertion
    • If sample rod was out in atmosphere overnight it may take several pumping cycles before it is ready to insert
  • Watch for green light to go on along with the orange
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Both lights will go out
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam
Added:
>
>

Aquiring a Beam

  • Open "HT" Menu (top left screen)
  • Select beam "on" *Filament current should be around 70%
 
Added:
>
>

Condenser Lens Alignment

  • Use "Brightness" knob to condense beam on function box(F)
  • press "Beam Shift" (F)
  • Right Click mouse to center beam
  • Spread Beam with "Brightness Knob (F)
  • If it does not spread evenly center with Condenser aperture x-y
  • condense beam with "Brightness" (F)
    • If beam is not centered, repeat
 
Added:
>
>

Alignment of DIffration Aperture

  • Select Diffraction button (F)
  • COndense beam with Brightness (F)
  • Insert Diffraction aperture
  • Center beam within aperture with diffraction aperture x-y align

Diffraction

  • Select Diffration button (F)
  • Use Align Adjust for Diffraction shift
  • PL Align

Low Dose

  • "Photo" menu
  • "MDS"
 

Specimen Removal

  • Make sure the beam is off
Changed:
<
<
  • Switch the toggle to "Air"
>
>
  • Switch the toggle to "Air" (G)
 
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
Changed:
<
<
  • Gently remove the specimen rod from the colum
>
>
  • Gently remove the specimen rod (G)
 
    • If the use of force is necessiary the SPC is not vented
Changed:
<
<
  • Set ALLOWTOPICVIEW =
>
>
Added:
>
>		* Set ALLOWTOPICVIEW =
 -- KdDerr - 13 Jun 2006

Revision 413 Jun 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Insertion

  • Line up pin on holder with the notched opening
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
    • You will hear a click
    • Orange light will go on
  • Flip toggle switch to "pump" (located below insertion point)
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • SPC value needs to be 40 or lower and read "Vac. Ready" before insertion
    • If sample rod was out in atmosphere overnight it may take several pumping cycles before it is ready to insert
  • Watch for green light to go on along with the orange
  • Turn holder slowly clockwise 1/8 turn to 1st stop
  • Slowly allow the vacuum to pull the holder in
  • Both lights will go out
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam

Specimen Removal

  • Make sure the beam is off
  • Switch the toggle to "Air"
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod from the colum
Changed:
<
<
>
>
    • If the use of force is necessiary the SPC is not vented
 
  • Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

Revision 313 Jun 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Changed:
<
<

Specimen Exchange

  • Line up pin on holder
>
>

Specimen Insertion

  • Line up pin on holder with the notched opening
 
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
Changed:
<
<
  • Flip toggle switch on goniometer to pump
>
>
    • You will hear a click
Added:
>
>
    • Orange light will go on
  • Flip toggle switch to "pump" (located below insertion point)
 
  • Moniter Vacuum conidtion:
Changed:
<
<
    • Go to "FR Panel" "moniter" "vacuum Conditions"
>
>
    • Go to "FR Panel" "moniter" "vacuum Conditions"
Added:
>
>
    • SPC value needs to be 40 or lower and read "Vac. Ready" before insertion
 
    • If sample rod was out in atmosphere overnight it may take several pumping cycles before it is ready to insert
Changed:
<
<
  • Watch for red light to go out
  • Turn holder slowly anti-clockwise to stop
>
>
  • Watch for green light to go on along with the orange
  • Turn holder slowly clockwise 1/8 turn to 1st stop
 
  • Slowly allow the vacuum to pull the holder in
Added:
>
>
  • Both lights will go out
  • Turn holder slowly clockwise 1/4 turn to 2nd stop
  • Slowly allow the vacuum to pull the holder in
  • Wait for green light to come on before turning on beam
 
Changed:
<
<

subsub level topic

>
>

Specimen Removal

  • Make sure the beam is off
Added:
>
>
  • Switch the toggle to "Air"
  • Moniter Vacuum conidtion:
    • Go to "FR Panel" "moniter" "vacuum Conditions"
  • Pull out specimen holder to 1st stop
  • Rotate anti-clockwise 1/4 turn
  • Pull out agian to the 2nd stop
  • Rotate anti-clockwise 1/8 turn
  • Wait for the airlock "SPC" to vent
  • Gently remove the specimen rod from the colum
 
  • Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

Revision 213 Jun 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Exchange

  • Line up pin on holder
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
  • Flip toggle switch on goniometer to pump
  • Moniter Vacuum conidtion:
Changed:
<
<
  • Go to "FR Panel" "moniter" "vacuum Conditions"

Watch for red light to go out Turn holder slowly anti-clockwise to stop

>
>
    • Go to "FR Panel" "moniter" "vacuum Conditions"
    • If sample rod was out in atmosphere overnight it may take several pumping cycles before it is ready to insert
  • Watch for red light to go out
  • Turn holder slowly anti-clockwise to stop
  • Slowly allow the vacuum to pull the holder in
Deleted:
<
<		Slowly allow the vacuum to pull the holder in
 

subsub level topic

  • Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

Revision 113 Jun 2006 - Main.KdDerr

 
META TOPICPARENT name="CryoEM"
Contents

JEOL 1230 Instructions

Specimen Exchange

  • Line up pin on holder
  • Insert specimen holder to first sample stop (marked line indicates correct depth)
  • Flip toggle switch on goniometer to pump
  • Moniter Vacuum conidtion:
  • Go to "FR Panel" "moniter" "vacuum Conditions"

Watch for red light to go out Turn holder slowly anti-clockwise to stop Slowly allow the vacuum to pull the holder in

subsub level topic

  • Set ALLOWTOPICVIEW =

-- KdDerr - 13 Jun 2006

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