Difference: Jeol3200Information (10 vs. 11)

Revision 1126 Apr 2015 - Main.BillRice

 
META TOPICPARENT name="CryoEM"

JEOL 3200 Microscope Parameters

wavelengths

  • 300kV: 0.01969 A

objective lenses

  • f= 4.4 mm
  • Cs = 4.1 mm
  • Chromatic aberration = 3.4 mm

Aperture Sizes

  • Condenser : 150 um, 70 um, 50 um, 20 um
  • Objective : 70 um, 50 um, 40 um, 20 um
  • Selected Area: 100 um, 50 um, 20 um, 10 um
  • Entrance Aperture : 120 um, 60 um, 40 um, 20 um

Energy Filter

  • Slit width at 10 eV : 60 mm on film
  • Slit width at 2 eV: 25 mm on film

Expected Values

  • Dark Current: 104 uA (12-5-11)
  • Emission Current: 179 uA (12-5-11)
  • Column / PiG1: 33 uA (12-5-11)
  • Camera / PiG3: 30 uA (12-5-11)
  • PEG1: 1.5 *10^-5 Pa (12-5-11)

Ice Thickness

The ice thickness was measured by punching a hole on the ice at 30 degrees (tilt) and measuring the length of the hole at -30 degrees (tilt). Using a collection semiangle of 45mrad, and assuming that the ice scatters as Carbon, the value that DM gives for the thickness is roughly half the actual thickness of the ice.
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Direct Electron Detector

  • Direct Electron DE20 detector installed 3/17/15
  • camera size: 5120 x 3840 pixels
  • pixel size: 6.4 um
  • active area: 33 mm * 25 mm
  • Survey Camera: 2048 x 2048 off-axis CCD
  • Survey Camera centered on scale bar visible on large screen
  • Sensor Cooling: Temperature when cooled should be 0C
  • Camera installed in film chamber, so mag ratio to film should be nearly 1.0
 

CCD information

  • Size of camera : 4096 x 4096 pixels
  • Pixel size: 15 um
  • Post-film mag : 1.40
  • Gain in SerialEM and Digital Micrograph : 29 counts / electron

Part number for replacement balloons

  • part #780 208 463
  • Used for filling liquid helium

CCD : Signal to noise ratio in comparison to film

Magnification Calibration: Jeol 3200FSC

  • Magnifications calibrated using 2160 cross grid using method described in SerialEM documentation
Film Magnification CCD Magnification Unbinned Pixel Size (A) 2X Binned Pixel Size (A) Angle of Tilt Axis Calibrated Film Magnification
LM 100 143 1046 2093 206.3  
LM 120 190 791 1583 212.5  
LM 150 248 606 1212 219.8  
LM 200 313 479 957 221.7  
LM 250 360 417 833 222.6  
LM 300 426 352 703 222.9  
LM 400 556 270 540 223.9  
LM 500 679 221 442 224.5  
LM 600 808 186 371 224.8  
LM 800 1,057 142 284 226.0  
LM 1,000 1,302 115 230 224.7  
LM 1,200 1,574 95.3 191 223.1  
LM 1,500 1,965 76.3 153 226.8  
LM 2,000 2,650 56.6 113 229.8  
LM 2,500 3,442 43.6 87.2 234.9  
LM 3,000 4,300 34.9 69.8 238.6  
MAG 2,000 2,750 54.5 109 155.6  
2,500 3,439 43.6 87.2 154.7  
3,000 4,160 36.1 72.1 154.9  
4,000 5,574 26.9 53.8 155.0  
5,000 6,794 22.1 44.2 125.7  
6,000 8,050 18.6 37.3 126.0  
8,000 10,841 13.8 27.7 126.2  
10,000 13,798 10.9 21.7 125.2  
12,000 16,598 9.04 18.1 125.8  
15,000 20,741 7.23 14.5 126.4  
20,000 26,806 5.60 11.2 126.1  
25,000 33,962 4.42 8.83 127.2  
30,000 40,859 3.67 7.34 127.3  
40,000 54,074 2.77 5.55 125.6  
50,000 68,165 2.20 4.40 126.0  
60,000 81,221 1.85 3.69 125.8 59,717
80,000 106,738 1.41 2.81 125.8  
100,000 132,895 1.13 2.26 125.8  
120,000 157,146 0.95 1.91    
* Overall CCD to film factor: 1.40

Defocus Offset

  • Set defocus to varying values between 500nm and 3000 nm
  • Focus at 200,000X, image at 50,000X on CCD
  • Defocus determined using ctffind3.
  • curve: calc_df = 0.76 * set_df + 36 nm
  • df offset curve may 9 2013:
    j3200_df_offset.png
  • Set ALLOWTOPICVIEW =

-- BillRice - 28 Sep 2007

META FILEATTACHMENT attachment="j3200_df_offset.png" attr="" comment="df offset curve may 9 2013" date="1368656112" name="j3200_df_offset.png" path="j3200_df_offset.png" size="3973" stream="j3200_df_offset.png" user="Main.BillRice" version="1"
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