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Electron Microscopy Protocols

Loading and Unloading the Gatan Rotational Holder into the JEOL 2010 Electron Microscope

4/11/01

 

Materials:

            JEOL 2010 Electron Microscope, Gatan Single Tilt Rotational Holder Model 650, motor drive control unit, mouse control, connecting wires with plug.

 

 

Procedure: Inserting the Gatan Rotational Holder

 

  1. Follow the diagrammed instructions for rotational holder insertion into the specimen chamber. Go to page 3 on the TV screen to observe the vacuum sequence. See diagram below:

 

Note that the rotational holder (RH) must be manually supported until the vacuum pull is strong enough to self-support in step #5.  It usually takes several minutes for the rotational holder with a glued grid/hex ring to pump down sufficiently until the green light comes on (step #4). Just be patient!

 

  1. Prepare to plug in the connecting cord into the handle of the RH. Note the contour of the plug jacket and the socket. See diagram below.

 

Note also that the plug has an outer sleeve that must be pulled back. Note the corresponding contour of the receptacle in the handle as well as the configuration of the prongs. Carefully connect the plug into the receptacle.

 

  1. If the insertion of the RH ever causes the vacuum to shut down, then suspect that the two black O-rings on the holder may need some greasing. In this case, remove the holder from the electron microscope. Then remove the two O-rings. Note that one is smaller than the other. With a Kim-wipe, clean each O-ring with 70% ethanol. Allow them to air dry. Then, smear a small amount of white grease on each ring. Wipe away excess with a Ross lens tissue. Next, re-attach the correct size O-ring to their positions on the RH. Finally, try re-inserting the RH into the electron microscope again.

 

  1. Once the specimen is in the electron microscope, set the MAG mode to LOW MAG, without any objective apertures in the way.

 

  1. Turn on the filament current to view the grid at 50X or 60X for several minutes. At the same time, exposure of the Pioloform coated grid to this low level electron beam current will serve to “condition” the entire grid. This “conditioning” should result in a more stabile Pioloform structure and consequently, less image drift.  While waiting, one can count the number of sections in the ribbon, also noting the location, orientation and disposition of the sections. One can also photograph the ribbon at LOW MAG.

 

 

Procedure: Removing the Gatan Rotational Holder

 

  1. Follow the diagrammed instructions for removing the holder from the column.

 

 

  1. Carefully unplug the connecting cord from the holder.

 

  1. Return rotational holder to stand.

Last Updated: 6/27/01