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Operating Procedures for TEM2 FEI Tecnai

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Operating Procedures for TEM2 FEI Tecnai Operating Procedures for TEM2 FEI Tecnai Note: Do not press the buttons on the TEM. This will turn the TEM off and will take hours to bring it back up. Please do not reboot the TEM computer. This will turn off the vacuum and the high tension. If you are having problems with the TEM, please find a SMIF staff member immediately. 1) Log usage into the SMIF web site. 2) Check to see if the camera boxes are on. a) The TIA camera box is the box behind the monitors. The switch will be lit green if on. b) The AMT camera box is on the floor under the Cryo holder. The green light above the switch should be on. c) If either of the cameras were off, please find a SMIF staff member to turn on. The cameras must cool down for 1hr before they can be used. 3) If needed, log into the TEM computer. User name is : TEM Users, Password is: tecnai 4) If needed, open the Tecnai User Interface and then open the TIA software. If the TIA software crashes, please find a SMIF staff member. 5) Place LN2 into the cold finger dewar. The LN2 needs to be topped off before each use. Turning on TEM Note: It is important to change the kV before ramping up the Heat to # when turning on the TEM. 1) There are tabs at the top left of the Tecnai software. Go to the Tune tab. Under the Control Pads box click on Fluorescent Background Light button. This turns on the lights on the control panels. 2) Press the Set up tab and look for the Vacuum box. 3) The Status bar (at top of the box) should read COL. VALVES. If it reads CRYO CYCLE or COL. COND, a cryo cycle is running and it has to complete before the TEM can be used. If there something other than the above or READY in the Status bar the TEM is not operational. Please contact a SMIF staff member. 4) Check the vacuum readings in the Vacuum box. The Gun should be less than 25 and the Column should be less that 15. If the values are great than this, please wait till they return to these values before continuing. 5) Check the High Tension setting (kV) and check to see if the High Tension button is yellow. If not yellow, press the button to turn on. a) If the button does not go yellow, check to see if the HT button on the TEM is lit. If it is not lit, press it to turn on. If it is lit, try pressing the High Tension button in the software again. If it still does not go yellow, find a SMIF staff member. 6) Change the kV if needed, using the pull down menu. 7) Adjust the Heat to # to the setting for filament saturation at your working kV (these are posted on the TEM). Press the enter button next to the Heat to #. The filament takes about 15mins to warm up from a Heat to # of 15 and 40mins if it was off. 8) Adjust Step # to the setting for the working kV (these are posted on the TEM). 9) Check the Filament button to see if it is yellow. If it is not, press it and it should turn yellow. 10) Adjust the Spot size to 3. 11) The emission current should read between 4-8 µA once the filament has reached the desired Heat to #. The emission current can be increased by changing the Step # one number at a time. 12) Press the Col Valves Closed button (button will turn grey). This will unblank the beam. TEM2 Operating Procedures Revision 6 Page 1 M. Plue 05/31/2017 13) Reduce the size of the beam with the Intensity knob on the left panel. Then bring the beam to the center of the screen with the track ball (also on the left panel). 14) Spread the beam with the Intensity knob so that it covers the entire viewing screen. Gain Reference with the TIA Camera Note: A gain reference should be acquired before imaging your samples. It needs to be completed at least once a day. 1) A sample should not be in the way of the beam. This includes films on a grid. 2) Adjust Exp time to 7-8 sec using the Intensity knob (Exp time is found on the bottom right of the TEM software). 3) Raise the viewing screen by pressing the R1 button on the right panel and place cover over viewing window. 4) Go to the CCD/TV tab and press the Preview button. 5) Press the arrow button at the top of the CCD/TV box. This will give you another menu screen. 6) Under the Bias/ Gain tab, press the All Gain button. 7) Click Yes in the text box on the image screen. The gain reference will automatically be acquired. 8) A text box will display on the image. When the gain reference is finished, please close the box. 9) Press R1 and lower the screen. 10) Place sample into the column (see instructions below). Gain Reference with the AMT Camera Note: A gain reference should be acquired before imaging your samples. It needs to be completed at least once a day. 1) A sample should not be in the way of the beam. This includes films on a grid. 2) Adjust exposure time to 3 sec using the Intensity knob. 3) Lower the viewing screen on the TEM by pressing the R1 button. 4) Press the scroll wheel and the right mouse button to toggle the mouse and keyboard to the AMT camera monitor. 5) Open the AmtV600 icon. 6) Press the Camera Out button to insert the AMT camera. 7) Press the Click for Live Image to view an image. 8) Select Corrections on the top menu bar. 9) Select Acquire a Background and then press the Proceed button. 10) When the gain reference is completed, remove camera by pressing the Camera In button. 11) Place sample into the column (see instructions below). Removing and Placing sample into TEM Note: Wear gloves when handling the sample holder. Never touch the gold part of the sample holder, even with gloves on. At the end of the session, replace the sample holder into the TEM. 1) Press the Set up tab and got to the Vacuum box. 2) Close the Column Valves by pressing the Col Valves Closed button (button will turn yellow). 3) Check the position of the sample holder. The x & y values are posted at the bottom right of the TEM software. If it is not at 0x & 0y, the stage should be returned to this position. TEM2 Operating Procedures Revision 6 Page 2 M. Plue 05/31/2017 a) To do this, go to the Stage tab and then go to the Stage box. Look for the arrow button. Press it and another Menu box will be displayed. b) Under the Control tab, find the XY button. Pressing the XY button will return the stage to 0x, 0y. 4) Return to the Set Up tab and note the vacuum values. 5) To remove the sample holder from the TEM, complete the following steps: a) Place one hand on blue/purple plate and grasp the handle of the sample holder with the other hand. Pull the sample holder slowly straight out of TEM until it stops. Do not let go or the sample holder will be pulled back into the TEM. This can damage the sample holder. b) Turn the holder half a turn clockwise and wait 10 seconds. c) Pull the sample holder straight out of the TEM. 6) Place the sample holder into the protective receptacle. Remove the end cap. 7) Raise the sample clamp with the pin. 8) Remove grid (if necessary) and place a new grid in the sample holder. 9) Lower the sample clamp with the pin and remove sample holder from receptacle. 10) To place the sample holder into the TEM, complete the following steps: a) Line the small post on the sample holder to 12 o’clock and the larger post on the handle to 6 o’clock. b) Slowly place the sample holder into the sample exchange until it stops. The red light will come on and the pump will start. c) Turn the sample holder ½ a turn clockwise. You will feel the sample holder slip into place. d) Select single tilt holder on the bottom menu bar and press the return button. e) Wait for the Status bar to change from Airlock to Col Valves. The red light on the stage will go off and the pump will stop. f) Rotate the sample holder counter-clockwise about a half turn. g) Insert the holder into the TEM slowly. Be careful not to let the holder be pulled into the TEM. 11) Check the Column pressure and Gun pressure (in the Vacuum box). If the Column pressure is greater than 15 log and/or the Gun pressure is greater than 25 log, wait until they return to 15 and 25 before opening the Column Valves. 12) Open the Column Valves by pressing the Col Valves Closed button (button will go gray). 13) If you do not do not see the beam, complete the following steps: a) Go down in magnification and move the stag (with joystick on right panel). b) If you still do not see the beam, turn the Intensity knob (on the left panel) both ways and move the beam (with the track ball on the left panel).
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