Return to Cryo EM web page at http://www.nysbc.org/facilities/CEM

To make a montage of your grid

1. Set magnification to 170X, maximum beam spread.
2. Change spot size 5 (with the console buttons, check Tecnai interface on left screen)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages
5. Go to Navigator Menu/Open, then Navigator/Montage/Super Montage/Setup Full Montage, Click OK
6. For the header of the montage file select all the available options
7. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
8. Go to Montage controls and click Start (to start the montage)
9. Save the montage as a (.st) stack file, double check the file extension (.st).
10. Save navigator file as .nav
11. When done, it will ask if you want to make a new map. ALWAYS SAY YES
12. Choose file-->close, otherwise you will not be able to change camera settings. This only closes the file for writing.

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to dose
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

To start next Tilt Series

1. Go back to 0 tilt (on tilt control menu)
2. Go to next area (predose if necessary)
3. If you've changed magnification, go back to desired mag
4. Do autofocus
5. Start new tilt series

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• When changing the binning value in Camera&Macro Ctrls after taking a full montage at 170x, don't forget to change the exposure time to 1 sec
• Set ALLOWTOPICVIEW =

-- MarienaSilvestry - 27 June 2011 | Modified from KdDerr r30 file

Return to Cryo EM web page at http://www.nysbc.org/facilities/CEM

To make a montage of your grid

1. Set magnification to 170X, maximum beam spread.
2. Change spot size 5 (with the console buttons, check Tecnai interface on left screen)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages
5. Go to Navigator Menu/Open, then Navigator/Montage/Super Montage/Setup Full Montage, Click OK
6. For the header of the montage file select all the available options
7. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
8. Go to Montage controls and click Start (to start the montage)
9. Save the montage as a (.st) stack file, double check the file extension (.st).
10. Save navigator file as .nav
11. When done, it will ask if you want to make a new map. ALWAYS SAY YES
12. Choose file-->close, otherwise you will not be able to change camera settings. This only closes the file for writing.

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box

• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

To start next Tilt Series

1. Go back to 0 tilt (on tilt control menu)
2. Go to next area (predose if necessary)
3. If you've changed magnification, go back to desired mag
4. Do autofocus
5. Start new tilt series

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• When changing the binning value in Camera&Macro Ctrls after taking a full montage at 170x, don't forget to change the exposure time to 1 sec
• Set ALLOWTOPICVIEW =

-- MarienaSilvestry - 27 June 2011 | Modified from KdDerr r30 file

Return to Cryo EM web page at http://www.nysbc.org/facilities/CEM

To make a montage of your grid

1. Set magnification to 170X, maximum beam spread.
2. Change spot size 5 (with the console buttons, check Tecnai interface on left screen)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages
5. Go to Navigator Menu/Open, then Navigator/Montage/Super Montage/Setup Full Montage, Click OK
6. For the header of the montage file select all the available options
7. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
8. Go to Montage controls and click Start (to start the montage)
9. Save the montage as a (.st) stack file, double check the file extension (.st).
10. Save navigator file as .nav
11. When done, it will ask if you want to make a new map. ALWAYS SAY YES
12. Choose file-->close, otherwise you will not be able to change camera settings. This only closes the file for writing.

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

To start next Tilt Series

1. Go back to 0 tilt (on tilt control menu)
2. Go to next area (predose if necessary)
3. If you've changed magnification, go back to desired mag
4. Do autofocus
5. Start new tilt series

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• When changing the binning value in Camera&Macro Ctrls after taking a full montage at 170x, don't forget to change the exposure time to 1 sec
• Set ALLOWTOPICVIEW =

Return to Cryo EM web page at http://www.nysbc.org/facilities/CEM

To make a montage of your grid

1. Set magnification to 170X, maximum beam spread.
2. Change spot size 5 (with the console buttons, check Tecnai interface on left screen)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages
5. Go to Navigator Menu/Open, then Navigator/Montage/Super Montage/Setup Full Montage, Click OK
6. For the header of the montage file select all the available options
7. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
8. Go to Montage controls and click Start (to start the montage)
9. Save the montage as a (.st) stack file, double check the file extension (.st).
10. Save navigator file as .nav
11. When done, it will ask if you want to make a new map. ALWAYS SAY YES
12. Choose file-->close, otherwise you will not be able to change camera settings. This only closes the file for writing.

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

To start next Tilt Series

1. Go back to 0 tilt (on tilt control menu)
2. Go to next area (predose if necessary)
3. If you've changed magnification, go back to desired mag
4. Do autofocus
5. Start new tilt series

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• When changing the binning value in Camera&Macro Ctrls after taking a full montage at 170x, don't forget to change the exposure time to 1 sec
• Set ALLOWTOPICVIEW =

Return to Cryo EM web page at http://www.nysbc.org/facilities/CEM

To make a montage of your grid

1. Set magnification to 170X, maximum beam spread.
2. Change spot size 5 (with the console buttons, check Tecnai interface on left screen)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages
5. Go to Navigator Menu/Open, then Navigator/Montage/Super Montage/Setup Full Montage, Click OK
6. For the header of the montage file select all the available options
7. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
8. Go to Montage controls and click Start (to start the montage)
9. Save the montage as a (.st) stack file, double check the file extension (.st).
10. Save navigator file as .nav
11. When done, it will ask if you want to make a new map. ALWAYS SAY YES
12. Choose file-->close, otherwise you will not be able to change camera settings. This only closes the file for writing.

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

To start next Tilt Series

1. Go back to 0 tilt (on tilt control menu)
2. Go to next area (predose if necessary)
3. If you've changed magnification, go back to desired mag
4. Do autofocus
5. Start new tilt series

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• When changing the binning value in Camera&Macro Ctrls after taking a full montage at 170x, don't forget to change the exposure time to 1 sec
• Set ALLOWTOPICVIEW =

Return to Cryo EM web page at http://www.nysbc.org/facilities/CEM

To make a montage of your grid

1. Set magnification to 170X, maximum beam spread.
2. Change spot size 5 (with the console buttons, check Tecnai interface on left screen)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages
5. Go to Navigator Menu/Open, then Navigator/Montage/Super Montage/Setup Full Montage, Click OK
6. For the header of the montage file select all the available options
7. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
8. Go to Montage controls and click Start (to start the montage)
9. Save the montage as a (.st) stack file, double check the file extension (.st).
10. Save navigator file as .nav
11. When done, it will ask if you want to make a new map. ALWAYS SAY YES
12. Choose file-->close, otherwise you will not be able to change camera settings. This only closes the file for writing.

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• When changing the binning value in Camera&Macro Ctrls after taking a full montage at 170x, don't forget to change the exposure time to 1 sec
• Set ALLOWTOPICVIEW =

Return to Cryo EM web page at http://www.nysbc.org/facilities/CEM

To make a montage of your grid

1. Set magnification to 170X, maximum beam spread.
2. Change spot size 5 (with the console buttons, check Tecnai interface on left screen)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages
5. Go to Navigator Menu/Open, then Navigator/Montage/Super Montage/Setup Full Montage, Click OK
6. For the header of the montage file select all the available options
7. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
8. Go to Montage controls and click Start (to start the montage)
9. Save the montage as a (.st) stack file, double check the file extension (.st).
10. Save navigator file as .nav
11. When done, it will ask if you want to make a new map. ALWAYS SAY YES
12. Choose file-->close, otherwise you will not be able to change camera settings. This only closes the file for writing.

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

Return to Cryo EM web page at http://www.nysbc.org/facilities/CEM

To make a montage of your grid

1. Set magnification to 170X, maximum beam spread.
2. Change spot size 5 (with the console buttons, check Tecnai interface on left screen)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages
5. Go to Navigator Menu/Open, then Navigator/Montage/Super Montage/Setup Full Montage, Click OK
6. For the header of the montage file select all the available options
7. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
8. Go to Montage controls and click Start (to start the montage)
9. Save the montage as a (.st) stack file, double check the file extension (.st).
10. Save navigator file as .nav
11. When done, it will ask if you want to make a new map. ALWAYS SAY YES
12. Choose file-->close, otherwise you will not be able to change camera settings. This only closes the file for writing.

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

Return to Cryo EM web page at http://www.nysbc.org/facilities/CEM

To make a montage of your grid

1. Set magnification to 170X, maximum beam spread.
2. Change spot size 5 (with the console buttons, check Tecnai interface on left screen)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages

1. For the header of the montage file select all the available options
2. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
3. Go to Montage controls and click Start (to start the montage)
4. Save the montage as a (.st) stack file, double check the file extension (.st).
5. Save navigator file as .nav
6. When done, it will ask if you want to make a new map. ALWAYS SAY YES
7. Choose file-->close, otherwise you will not be able to change camera settings. This only closes the file for writing.

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

Return to Cryo EM web page at http://www.nysbc.org/facilities/CEM

To make a montage of your grid

1. Set magnification to 170X, maximum beam spread.
2. Change spot size 5 (with the console buttons, check Tecnai interface on left screen)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages

1. For the header of the montage file select all the available options
2. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
3. Go to Montage controls and click Start (to start the montage)
4. Save the montage as a (.st) stack file, double check the file extension (.st).
5. Save navigator file as .nav
6. When done, it will ask if you want to make a new map. ALWAYS SAY YES
7. Choose file-->close, otherwise you will not be able to change camera settings. This only closes the file for writing.

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

Return to Cryo EM web page at http://www.nysbc.org/facilities/CEM

To make a montage of your grid

1. Set magnification to 170X, maximum beam spread.
2. Change spot size 5 (with the console buttons, check Tecnai interface on left screen)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages

1. For the header of the montage file select all the available options
2. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
3. Go to Montage controls and click Start (to start the montage)
4. Save the montage as a (.st) stack file, double check the file extension (.st).
5. Save navigator file as .nav
6. When done, it will ask if you want to make a new map. ALWAYS SAY YES
7. Choose file-->close, otherwise you will not be able to change camera settings. This only closes the file for writing.

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

Return to Cryo EM web page at http://www.nysbc.org/facilities/CEM

To make a montage of your grid

1. Set magnification to 170X, maximum beam spread.
2. Change spot size 5 (with the console buttons, check Tecnai interface on left screen)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages
5. Go to Navigator Menu/Open, then Menu/Montage/Super Montage/Setup Full Montage, Click OK
6. For the header of the montage file select all the available options
7. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.

1. Save the montage as a (.st) stack file, double check the file extension (.st).
2. Save navigator file as .nav
3. When done, it will ask if you want to make a new map. ALWAYS SAY YES
4. Choose file-->close, otherwise you will not be able to change camera settings. This only closes the file for writing.

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

Return to Cryo EM web page at http://www.nysbc.org/facilities/CEM

To make a montage of your grid

1. Set magnification to 170X, maximum beam spread.
2. Change spot size 5 (with the console buttons, check Tecnai interface on left screen)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages
5. Go to Navigator Menu/Open, then Menu/Montage/Super Montage/Setup Full Montage, Click OK
6. For the header of the montage file select all the available options
7. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.

1. Save the montage as a (.st) stack file, double check the file extension (.st).
2. Save navigator file as .nav
3. When done, it will ask if you want to make a new map. ALWAYS SAY YES
4. Choose file-->close, otherwise you will not be able to change camera settings. This only closes the file for writing.

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

Return to Cryo EM web page at http://www.nysbc.org/facilities/CEM

To make a montage of your grid

1. Change spot size 5 (with the console buttons, check Tecnai interface on left screen)
2. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
3. Make sure to increase registration number if you intend to transfer points from previous montages
4. Go to Navigator Menu/Open, then Menu/Montage/Super Montage/Setup Full Montage, Click OK
5. For the header of the montage file select all the available options
6. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
7. Go to Montage controls windows and click Start (to start the montage)
8. Save the montage as a (.st) stack file, double check the file extension (.st).
9. Save navigator file as .nav
10. When done, it will ask if you want to make a new map. ALWAYS SAY YES
11. Choose file-->close, otherwise you will not be able to change camera settings. This only closes the file for writing.

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

Return to Cryo EM web page at http://www.nysbc.org/facilities/CEM

To make a montage of your grid

1. Set magnification to 170X
2. Change spot size 5 (with the console buttons, check Tecnai interface on left screen)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages

1. For the header of the montage file select all the available options
2. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.

1. Save navigator file as .nav
2. When done, it will ask if you want to make a new map. ALWAYS SAY YES
3. Choose file-->close, otherwise you will not be able to change camera settings. This only closes the file for writing.

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

Return to Cryo EM web page at http://www.nysbc.org/facilities/CEM

To make a montage of your grid

1. Set magnification to 170X

1. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
2. Make sure to increase registration number if you intend to transfer points from previous montages

1. For the header of the montage file select all the available options
2. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
3. Go to Montage controls/Start (to start the montage)
4. Save the montage as a (.st) stack file.
5. Save navigator file as .nav
6. When done, it will ask if you want to make a new map. ALWAYS SAY YES
7. Choose file-->close, otherwise you will not be able to change camera settings. This only closes the file for writing.

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

To make a montage of your grid

1. Set magnification to 170X
2. Change spot size 5 (with the console buttons)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages
5. Go to Navigator menu/Open/Montage/Super Montage/Setupfull montage Click OK
6. For the header of the montage file select all the available options
7. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
8. Go to Montage controls/Start (to start the montage)
9. Save the montage as a (.st) stack file.
10. Save navigator file as .nav
11. When done, it will ask if you want to make a new map. ALWAYS SAY YES
12. Choose file-->close, otherwise you will not be able to change camera settings. This only closes the file for writing.

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

To make a montage of your grid

1. Set magnification to 170X
2. Change spot size 5 (with the console buttons)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages
5. Go to Navigator menu/Open/Montage/Super Montage/Setupfull montage Click OK
6. For the header of the montage file select all the available options
7. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
8. Go to Montage controls/Start (to start the montage)
9. Save the montage as a (.st) stack file.
10. Save navigator file as .nav
11. When done, it will ask if you want to make a new map. ALWAYS SAY YES
12. Choose file-->close, otherwise you will not be able to change camera settings. This only closes the file for writing.

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

To make a montage of your grid

1. Set magnification to 170X
2. Change spot size 5 (with the console buttons)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages
5. Go to Navigator menu/Open/Montage/Super Montage/Setupfull montage Click OK
6. For the header of the montage file select all the available options
7. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
8. Go to Montage controls/Start (to start the montage)
9. Save the montage as a (.st) stack file.
10. Save navigator file as .nav
11. When done, it will ask if you want to make a new map. ALWAYS SAY YES

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

To make a montage of your grid

1. Set magnification to 170X
2. Change spot size 5 (with the console buttons)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages
5. Go to Navigator menu/Open/Montage/Super Montage/Setupfull montage Click OK
6. For the header of the montage file select all the available options
7. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
8. Go to Montage controls/Start (to start the montage)
9. Save the montage as a (.st) stack file.
10. Save navigator file as .nav

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To Gain with Camera

• Go to Camera/Gain Ref Policy(default4900)

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

To make a montage of your grid

1. Set magnification to 170X
2. Change spot size 5 (with the console buttons)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages
5. Go to Navigator menu/Open/Montage/Super Montage/Setupfull montage Click OK
6. For the header of the montage file select all the available options
7. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
8. Go to Montage controls/Start (to start the montage)
9. Save the montage as a (.st) stack file.
10. Save navigator file as .nav

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

To make a montage of your grid

1. Set magnification to 170X
2. Change spot size 5 (with the console buttons)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages
5. Go to Navigator menu/Open/Montage/Super Montage/Setupfull montage Click OK
6. For the header of the montage file select all the available options
7. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
8. Go to Montage controls/Start (to start the montage)
9. Save the montage as a (.st) stack file.
10. Save navigator file as .nav

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Predosing Sample

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

To make a montage of your grid

1. Set magnification to 170X
2. Change spot size 5 (with the console buttons)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages
5. Go to Navigator menu/Open/Montage/Super Montage/Setupfull montage Click OK
6. For the header of the montage file select all the available options
7. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
8. Go to Montage controls/Start (to start the montage)
9. Save the montage as a (.st) stack file.
10. Save navigator file as .nav

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

To make a montage of your grid

1. Set magnification to 170X
2. Change spot size 5 (with the console buttons)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages
5. Go to Navigator menu/Open/Montage/Super Montage/Setupfull montage Click OK
6. For the header of the montage file select all the available options
7. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
8. Go to Montage controls/Start (to start the montage)
9. Save the montage as a (.st) stack file.
10. Save navigator file as .nav

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

• Go to 150 micron Condenser 2 aperture #4
• Obtain a new Gain Reference to calibrate dose
  • Take record image use ctl + i to get stats
  • Camera/Prepare Gain Ref...
  • Enter mean value in target dose box
• Add stage positions of areas to does
• Check Acquire box for each point
• Navigator/Acquire at points...
• Select Macro 4 to dose 2000e-
• Select Close Column Valves when done

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

To make a montage of your grid

1. Set magnification to 170X
2. Change spot size 5 (with the console buttons)
3. Go to Camera&Macro Controls Setup/Record tab Bining: 8 Exposure: 0.1 second
4. Make sure to increase registration number if you intend to transfer points from previous montages
5. Go to Navigator menu/Open/Montage/Super Montage/Setupfull montage Click OK
6. For the header of the montage file select all the available options
7. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
8. Go to Montage controls/Start (to start the montage)
9. Save the montage as a (.st) stack file.
10. Save navigator file as .nav

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

To make a montage of your grid

1. Set magnification to 170X

1. For the header of the montage file select all the available options
2. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
3. Go to Montage controls/Start (to start the montage)
4. Save the montage as a (.st) stack file.
5. Save navigator file as .nav

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

To make a montage of your grid

1. Set magnification to 170X
2. Change spot size 4 (with the console buttons)
3. Go to Setup menu/Record Bining: 8 Exposure: 0.1 second
4. Go to Navigator menu/Open/Montage/Super Montage/Setupfull montage Click OK
5. For the header of the montage file select all the available options
6. 10 % overlap approximately 11x11 pieces. Larger files will crash in loading.
7. Go to Montage controls/Start (to start the montage)
8. Save the montage as a (.st) stack file.
9. Save navigator file as .nav

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Aligning rotated montage in Navigator

1. Increase registration
2. Collect new montage with identical parameters
3. Select a minimum of 3 points on both maps
   • Magnifying the image when picking points is helpful to make the marked points more accurate.
4. Go to each point and check Registration Point box on top of Navigator panel.
   • 1R1 = 2R1, 1R2 = 2R2, etc
5. Go to Navigator on pull down menu and select Transform Items

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

To make a montage of your grid

1. Set magnification to 170X
2. Change spot size 4 (with the console buttons)
3. Go to Setup menu/Record Bining: 8 Exposure: 0.1 second
4. Go to Navigator menu/Open/Montage/Super Montage/Setupfull montage Click OK
5. For the header of the montage file select all the available options

1. Go to Montage controls/Start (to start the montage)
2. Save the montage as a (.st) stack file.
3. Save navigator file as .nav

Low Dose Montage

1. Go to exposure mode in Tecnai Low Dose
2. Open SerialEM
   • Once SerialEM is open do not use Tecnai interface to control microscope
3. See above instructions
4. After collecting montage pick points, move to area and save point in the Tecnai Specimen Position feature
5. Close SerialEM
6. Resume normal Low Dose operations

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

To make a montage of your grid

1. Set magnification to 170X
2. Change spot size 4 (with the console buttons)
3. Go to Setup menu/Record Bining: 8 Exposure: 0.1 second
4. Go to Navigator menu/Open/Montage/Super Montage/Setupfull montage Click OK
5. For the header of the montage file select all the available options
6. Go to Montage controls/Start (to start the montage)
7. Save the montage as a (.st) stack file.
8. Save navigator file as .nav

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

To make a montage of your grid

1. Set magnification to 170X
2. Change spot size 4 (with the console buttons)
3. Go to Setup menu/Record Bining: 8 Exposure: 0.1 second
4. Go to Navigator menu/Open/Montage/Super Montage/Setupfull montage Click OK
5. For the header of the montage file select all the available options
6. Go to Montage controls/Start (to start the montage)
7. Save the montage as a (.st) stack file.
8. Save navigator file as .nav

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

To make a montage of your grid

1. Set magnification to 170X
2. Change spot size 4 (with the console buttons)
3. Go to Setup menu/Record Bining: 8 Exposure: 0.1 second
4. Go to Navigator menu/Open/Montage/Super Montage/Setupfull montage Click OK
5. For the header of the montage file select all the available options
6. Go to Montage controls/Start (to start the montage)
7. Save the montage as a (.st) stack file.
8. Save navigator file as .nav

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

To make a montage of your grid

1. Set magnification to 170X
2. Change spot size 4 (with the console buttons)
3. Go to Setup menu/Record Bining: 8 Exposure: 0.1 second
4. Go to Navigator menu/Open/Montage/Super Montage/Setupfull montage Click OK
5. For the header of the montage file select all the available options
6. Go to Montage controls/Start (to start the montage)
7. Save the montage as a (.st) stack file.
8. Save navigator file as .nav

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

To make a montage of your grid

1. Set magnification to 170X
2. Change spot size 4 (with the console buttons)
3. Go to Setup menu/Record Bining: 8 Exposure: 0.1 second
4. Go to Navigator menu/Open/Montage/Super Montage/Setupfull montage Click OK
5. For the header of the montage file select all the available options
6. Go to Montage controls/Start (to start the montage)
7. Save the montage as a (.st) stack file.
8. Save navigator file as .nav

Tomogram settings

• File/close (to close montage image stack)
• Go to Setup menu/Record Bining: 2 Exposure: 1 second

To start a Tilt Series

1. Go to Tilt Series menu/Setup/Start
2. Select desired parameters

To finish a Tilt Series

1. After the tilt series is done go to Tilt Series menu/Terminate
2. Select "Yes" to close image stack
3. Select "No" to save 90 degrees rotated image in Buffer H

Useful Tip

• If you want to go to a particular slice of your stack file go to the File menu/Read/select file/select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

To make a montage of your grid

1. Set magnification to 170X
2. Change spot size 4 (with the console buttons)

1. For the header of the montage file select all the available options

To start a Tilt Series

1. Select desired parameters

To finish a Tilt Series

Useful Tip

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009

When you start with the TF20

1- Check that the IGP1 value is below 15 then is ready to use 2- Drop the screen with some of the console buttons (check Serial EM for the function of each of the buttons)

To make a montage of your grid

1- Set magnification to 170X 2- Change spot size 4 (with the console buttons) 3- Go to Setup menu  Record  Bin 8  exposure 0.1 second 4- Go to Navigator menu  Open  Montage/Super Montage  Setup  full montage  Click OK 5- For the header of the montage file select all the available options 6- Go to Montage controls  Start (to start the montage) 7- Save the montage as a stack file.

For tomography select a bining of 2 and a 1 second exposure

To start a Tilt Series

1- Go to Tilt Series menu  Setup/Start 2- Select desired parameters

To finish a Tilt Series

1- After the tilt series is done go to Tilt Series menu  Terminate 2- Select Yes in the message on your screen 3- Select No to save 90 degrees rotated

Useful Tip

If you want to go to a particular slice of your stack file go to the File menu  Read  select file  select slice

F20 Defocus table for common magnifications

• Set ALLOWTOPICVIEW =

-- KdDerr - 02 Oct 2009