2.1. Log on to the TUMI system (you cannot proceed further until this is done).

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1 FEI DB235 ex-situ lift out TEM sample preparation procedure Nicholas G Rudawski ngr@ufledu (805) Last updated: 06/19/15 DISCLAIMER: this procedure describes one specific method for preparing ex-situ lift-out TEM samples via FIB It is not by any means universal, for there is no one correct way to make a sample Depending on the material, type of analysis needed, and amount of area needed for imaging, significant deviations from this guide may be necessary You should always check the literature to see what, if anything, has been published regarding FIB-based TEM sample preparation of your materials and/or analysis needs That being said, this method has been successfully applied to a wide range of materials where high-resolution TEM imaging over a relatively small analysis area is needed 1 Preliminary sample preparation 11 Prior to FIB processing, samples may need to be coated with a conductive layer of sufficient thickness to prevent surface damage during the initial stages of I-beam imaging and protective Pt layer deposition If surface preservation is not needed and the specimen is conductive, this is not needed However, this is needed if the specimen is insulating, even if surface preservation is not required because it will be impossible to get a stable I-beam image of an insulating specimen otherwise A layer >200 nm-thick of conductive C (three coatings from the MAIC evaporator) or Cr are known to work well for this application Other materials may also work well, but Au-Pd is not recommended for a protective conductive layer since it is known to become porous upon exposure to the I-beam 2 Sample loading 21 Log on to the TUMI system (you cannot proceed further until this is done) 22 The FIB software (xp) should already be open and running Login with your username and password (you will receive this at the start of training) 23 On the far upper right corner of the software, enter the key panel by clicking on the icon 24 Under the Vacuum control click Vent to start venting 25 It will take 2 3 min for the chamber to vent You will know the chamber is vented when the door can be pulled open with little resistance 1

2 26 Insert the pin end of the stub into the stage and firmly tighten the setscrew Use the metal dog-ear and adjust the Z control knob on the outside of the door until the top of the sample is just under the bottom edge of the dog-ear 27 Remove the dog-ear from the stage, close the chamber door and click Pump ; be sure to apply light pressure to the door during the initial pump down 3 During evacuation 31 In the Confirm holder dialogue box that pops up after starting pumping, under Select Holder select Stub Holder if using a single stub or Multi- Stub Holder if using a multi-stub holder and select OK 32 While the chamber is pumping down, if Pt deposition will be needed during your session, enter the menu, go to GIS Heating and click On next to PT GIS 1 to start heating the Pt The Status light will change from blue to red when the Pt is ready to be used 33 Along the top menu bar, select E-Mag and make sure Mags Coupled is checked from the dropdown menu 2

3 34 Along the row of icons below the menu bar, select and to active the I-beam 35 Along the top menu bar, select Stage and select Zero Beam Shift from the dropdown menu to reset the beam shift for the I-beam 36 Along the row of icons below the menu bar, select and to activate the E-beam 37 Along the top menu bar, select Stage and select Zero Beam Shift from the dropdown menu to reset the beam shift for the E-beam 3

4 38 In the menu, go down to the Status and check that Vacuum Status reads Vac OK AND Chamber Pressure is < 10E-4 mbar and stable 4 Turning on the beams and finding an area of interest 41 In the menu under System, select Beams On to turn on the beams 42 Under Status, check that the Ion Column HV is reading 30 kv, Ion Emission Current is ~25 µa, and E Column HV and E Spotsize are reading 50 kv and 3, respectively (appropriate settings for general E- beam imaging) This will usually take ~30 sec 43 Make sure and are selected to active the E-beam 44 Select to start live imaging with the E-beam 45 Use the joystick and move around until you find a general area of interest where you wish to make a sample (alternatively, you can double click on a point in the image and the stage will move to center that point in the image) 5 Setting eucentric height 51 Enter the Swiss army knife panel by selecting the icon to the right of the icon 52 Use the magnification selector knob on the desktop control panel to increase the magnification to ~1 k 4

5 53 Find and center in the image a flat recognizable feature on the sample surface near the area of interest 54 Select (stage cannot be tilted unless this is done) 55 In the bottom of the panel, find the stage tilt T and input a value of 5 The image will shift either up or down (probably down) as stage is tilted 56 Use the Z control knob on the chamber door and adjust until the feature is once again centered 57 Repeat the previous 3 steps inputting T = 10, 20, 35, and Return the stage tilt to T = 0 59 Find and center the feature in the image 510 Increase the magnification to ~10k (this will be the magnification used for much of rest of the sample prep procedure) and focus the image using the desktop control panel 511 Tilt to T = 52 and adjust Z to return the feature to the center of the image 512 Return the stage tilt to T = Find and center the feature in the image (it shouldn t have moved much), and focus the image using the desktop control panel 514 Tilt to T = 52 and adjust Z to return the feature to center of the image (this should need little, if any, adjustment at this point) 515 Leave the stage tilt set to T = 52 6 Linking the beams 61 Make sure the stage tilt is set to T = In the menu bar under I-beam, select 30 pa and then select and to active the I-beam for imaging 5

6 63 On the right panel, select Scan Rotation and input 180 ; this rotates the I-beam image 180 relative to E-beam image and preserves the top-down orientation when the stage is tilted (NOTE: some users may opt to leave the I-beam scan rotation at 0 ; this is a matter of stylistic preference, but I recommend leaving it at 180 to start with) 64 Select the auto contrast/brightness icon (or press F9) to adjust the image for optimal viewing 65 Use the shift knobs (DO NOT translate the stage!) on the desktop control panel to move the I-beam image so the center of the image is coincident with the same point in the E-beam image (where you want to make your TEM sample); this links the beams so that the E-beam and I-beam image the same area on the sample If you properly set eucentric height, the need to shift the I-beam image should be minimal 66 If needed, you can translate the stage left or right by a small amount to move to a different area (perhaps a cleaner one); however, do not translate the stage up or down or you will no longer be at eucentric height 67 Select to stop live I-beam imaging 7 Protective Pt strip deposition 71 Make sure the stage tilt is set to T = 52 ; if you start this procedure at T = 0, your sample won t turn out right! 72 In the menu bar under I-beam, select a 300 pa setting 6

7 73 Under Gas Injection click In next to PT GIS 1 74 Along the row of milling patterns icons, select the filled box pattern icon and give it X, Y, and Z dimensions of 10, 10, and 10 µm (NOTE: these are only suggestions; you can adjust the dimensions to whatever is required for your samples) 8 75 Under Material File select pt_temmtr of the box should be green) (the outline 76 Perform auto contrast/brightness by selecting every time the I-beam current is changed) 77 Select 78 Select the arrow icon to position the box in the desired location for Pt deposition (try to stay near the center of the image) 79 Select the patterning icon (this should be done to take a single I-beam image to start deposition Milling deep trenches 7

8 81 Select Out next to PT GIS 1 to retract the Pt needle 82 Change the I-beam current to 5000 pa and perform auto contrast/brightness 83 Take a single I-beam image 84 Change Material File to simtr should change to yellow) 85 Erase the pattern by selecting the eraser icon 86 Select the regular cross-section pattern icon and click/drag/release on the image to draw the pattern Set the X, Y, and Z dimensions to 12, 4, and 20 µm (NOTE: for Si, milling trenches with Z = 20 µm will actually produce trenches ~60 µm deep, just FYI) 87 Position this pattern centered and below the Pt strip 88 Take a single I-beam image to see if any drift occurred and adjust the pattern position if necessary and then start the pattern 89 Select Edit under Patterning Advanced Features dialogue box 8 (the pattern outline and input 180 for rotation in the

9 810 Take a single I-beam image and then position the pattern centered and above the Pt strip NOTE: whenever a regular cross-section pattern is executed below (above) the Pt strip, the pattern rotation should be set to 0 (180 ) Take another single I-beam image to make sure the image is stable (adjust the pattern position, if necessary) and then start the pattern Rough thinning cuts 91 Change the I-beam current to 1000 pa, perform auto contrast/brightness, and take a single I-beam image 92 Select the arrow icon, click in the middle of pattern, and change the Y and Z dimensions to 05 and 10 µm 93 Change the pattern rotation to 0 94 Take another single I-beam image to make sure the image is stable, position the pattern centered and below the Pt strip, and start patterning 95 Change the pattern rotation to 180, then take a single I-beam image 9

10 10 96 Position the pattern centered and above the Pt strip and start the pattern 97 Perform the previous two milling steps (alternating) until the Pt layer is ~075 µm-wide (Y dimension) Undercutting 101 Change the stage tilt to T = When the stage is done tilting, take a single I-beam image auto contrast/brightness ; perform if necessary 103 Erase the pattern and then select the filled box pattern icon Draw one pattern with X, Y, and Z dimensions 12, 07, and 10 and two other patterns with X, Y, and Z dimensions of 07, 30, and 10 µm 104 Position the first pattern centered and ~2/3 of the distance from the sample surface to the bottom of the trench; position the other two patterns along the outer edges of the sample making sure the outer patterns are slightly below the surface of the sample 10

11 Take a single I-beam image and adjust the patterns if any drift occurred (NOTE: it may take a few seconds for the drift to settle after stage tilting), and then start the patterns 106 You can check the progress of the undercut by intermittently taking single E-beam images during the milling The sample will look similar to the following when the undercut has gone completely through Basically, you need to verify the U-shaped undercut has gone completely through the specimen (otherwise, you won t be able to lift it out!) Fine thinning cuts 111 The next series of steps alternates executing patterns from the top and bottom of the sample with stage tilts of 54 and 50 and pattern rotations of 180 and 0, respectively; this helps to preserve the protective Pt strip while creating a more uniformly-thick sample 112 Increase magnification to 15k for the rest of the procedure (optional, but helpful) 113 Tilt the stage to T = Erase the solid box patterns used for the undercut, draw a regular cross-section pattern, and set the X, Y, and Z pattern dimensions to 12, 025, and 05 µm (make sure the pattern rotation is set to 180 ) 115 Reduce the I-beam current to 300 pa, perform auto contrast/brightness, and take a single I-beam image 116 Position the pattern along the top edge of the sample and start the pattern Occasionally take E-beam images during milling to make sure the Pt strip is still intact 11

12 117 Tilt the stage to T = 50 and change the pattern rotation back to Take single I-beam images to verify the sample isn t drifting, then position the pattern below the bottom edge of the sample and start the pattern Occasionally take E-beam images during milling to make sure the Pt strip is still intact The sample should be ~03 µm thick after this step 119 Tilt the stage to T = 54, reduce the I-beam current to 100 pa, perform auto contrast/brightness, and take a single I-beam image 1110 Reduce the X dimension of the pattern to 60 µm and change the pattern rotation to Take a single I-beam image to make sure the sample isn t drifting, position the pattern centered and over the top edge of the sample and start the pattern During milling, frequently take E-beam images to make sure the Pt strip is still intact 12

13 1112 Tilt the stage to T = 50 and change the pattern rotation to Take single I-beam images to ensure the sample isn t drifting, position the pattern centered and over the bottom edge of the sample, and start milling During milling, frequently take E-beam images to make sure the Pt strip is still intact 1114 Repeat the previous 2 milling steps (alternatively) until the sample is thin enough for TEM; the sample is usually thin enough for TEM when the Pt layer decreases in height and becomes very bright The sample may now be freed for lift-out or given additional low kv cleaning to further thin it and reduce the damage layer 12 Low kv cleaning (a good idea for HR-TEM imaging) 13

14 121 To perform low kv cleaning, change the I-beam to Low kv: 300 pa 500 kv This will ramp the I-beam voltage down to 5 kv over a few seconds 122 Select and to activate the E-beam and select to start live imaging; the magnification will now be ~100 as a result of switching to the low kv I-beam setting (the magnifications will still be coupled) 123 Increase the magnification back to ~15k and select to stop live E- beam imaging 124 Select and to activate the I-beam, perform auto contrast/brightness, and take a single I-beam image ; due to the reduced I-beam voltage, the I-beam image resolution will not be as good as when the voltage was set to 30 kv 125 Tilt the stage to 57 and take a single I-beam image 126 Draw a filled box with X, Y, and Z dimensions of 60, 04, and 05 µm (the pattern rotation does not need to be adjusted here) 127 Take single I-beam images to make sure the sample is not drifting, and position the pattern so the bottom edge of the pattern is over the top edge of the sample; this will result in most of the sample being covered by the milling pattern (don t freak out, this is what you want to do) 128 Start the pattern and take frequent E-beam images during milling to make sure the sample thins a little further but the Pt strip remains intact 129 Tilt the stage to 47 and take single I-beam images to make sure the sample is not drifting 14

15 1210 Position the pattern so the top edge of the box is over the bottom edge of the sample; again this will result in most of the sample being covered by the milling pattern 1211 Start the pattern and take frequent E-beam images during milling to make sure the sample thins a little further but the Pt strip remains intact 1212 If the sample was thin enough after thinning with the I-beam at 30 kv, one cleaning step per side should be sufficient, but the previous two steps can be repeated, if needed 13 Release cuts (shown here with the low kv I-beam used for final thinning) 131 Leave the I-beam at the last setting used for milling the sample; for example, if the low kv setting was used as the last step, the low kv Ibeam setting should be used for doing the release cuts 132 Tilt the stage to T = 52 ; take single I-beam images sample is not drifting 133 Draw a filled box pattern 07, 10, and 20 µm 134 Position the pattern over one attached end of the sample and start milling ; take frequent E-beam images during milling to check if the end to make sure the with X, Y, and Z dimensions of the pattern to has been freed; stop the milling pattern immediately when the end has been released If the end wasn t freed after this step, repeat this step again 15

16 135 Repeat the previous step for the remaining attached end of the sample; make sure to take frequent E-beam images during milling to check if the sample has been freed; the sample should appear to have moved in the E-beam image after being freed Stop the milling pattern immediately when the sample has been freed 136 Change the I-beam back to the 30 pa 30 kv setting DO NOT do any more imaging of the specimen with the I-beam or the specimen will unnecessarily acquire further damage 137 The sample is now ready for ex-situ lifting out using the micro-manipulator system To make another sample in another location, first tilt the stage back to T = 0, select the E-beam and enter live imaging, and find your new area of interest You will have to reset eucentric height and re-link the beams as described earlier before making another sample 14 Finishing the session 141 Make sure the I-beam is set to the 30 pa 30 kv setting (DO NOT leave the I-beam at the Low kv setting when finishing your session) 142 Tilt the stage back to T = Enter the key menu and turn off the Pt heating (the On button should turn gray) 16

17 144 Turn off the beams; the I-Beam HV and E-beam HV will be 1 and 0 kv when the beams have been turned off, respectively (takes a few seconds to complete) 145 Select to freeze the live image; (DO NOT continue live imaging with the beams off) 146 When the beams are off, select Vent under the Vacuum control and agree to the dialogue box that pops up asking if you want to proceed with venting 147 After venting has started, select Stage from the main menu bar and select Initialize Stage from the dropdown menu Select Ok from the dialogue box that comes up 148 Once the chamber is vented, remove your sample, close the chamber door, and click Pump under the Vacuum control 17

18 149 Log off the TUMI system Appendix A: shorthand description of above procedure (steps 4 13 only) 1 Find and align area of interest using E-beam 2 Set eucentric height 3 Tilt stage to 52 and link beams with I-beam scan rotation set to180 4 Deposit Pt layer with 300 pa I-beam 5 Cut trenches on top and bottom side of Pt layer with 5000 pa I-beam 6 Performing rough thinning cuts on top and bottom side of Pt layer with 1000 pa I- beam until Pt layer is ~075 µm wide 7 Tilt to 7 and perform undercut with 1000 pa I-beam 8 Reduce I-beam current to 300 pa and perform alternating thinning cuts on top and bottom sides of the Pt layer with stage tilts 54 and 50 until sample is ~03 µm wide 9 Reduce I-beam current to 100 pa and perform alternating thinning cuts on top and bottom sides of the Pt layer with stage tilts 54 and 50 until sample is (or is nearly) electron transparent; take frequent E-beam images during milling to make sure Pt layer is intact if surface preservation is needed 10 If desired, I-beam voltage can be reduced to low kv setting and alternating cleaning steps performed from top and bottom sides of sample at stage tilts of 57 and 47 to further thin the sample and reduce the damage layer 11 When the specimen is finished, perform release cuts using the last used I-beam setting; take frequent E-beam images and stop milling as soon as specimen is free 12 Return I-beam to 30 pa 30 kv setting; DO NOT perform anymore I-beam imaging of the specimen 18

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