1 Nanosurf EasyScan 2 Atomic Force Microscope 2 Standard Operating Procedures. 3 4 Instrument set-up 5 Assemble and collect all the hardware in the box together 6 1. Connect cable A between camera and the controller video port. 7 2. Connect cable B between microscope and the front port of the controller. 8 3. Connect cable C between computer and the back port of the controller. 9 Mount the probe 10 4. Lay the probe holder on the bench upside down with the grove facing up. 11 12 5. Apply a downward pressure to the holder to open up the spring clip. With the spring clip open, insert the AFM chip into the groove. Release the holder to allow the spring clip hold the AFM chip. 13 14 6. Slightly release the spring clip and push the AFM chip with tweezers to center it and make a good contact with the groove. 15 Mount the probe holder 16 17 7. Insert the probe holder into the AFM head from top at an angle. Push gently forward, the lower the probe holder. 18 19 8. Check the position of the probe holder. The 3 precision ball mounts inside the AFM head should mate with the probe holders underside groves. 20 21 9. Place the AFM head on the sample stage and make sure the application is “off”, Place the assembly on the vibration absorber stage. 22 10. Roughly level the sample stage with 3 screws inside the head. 23 24 Sample Mounting and Manual Approach 25 1. Put on clean, disposable gloves. 26 27 2. Mount the scan head onto the sample stage, then place assembly on vibration isolation stage. Make sure application is off. 28 29 30 3. Rough level the scan head by turning the three edge screws until bubble on top of head is roughly centered within circle. Turning screws clockwise will raise the head; counterclockwise with lower it. Maintain enough height to prevent contact with sample holder during sample loading, 3-5 cm. 31 32 33 4. Secure the (dust free) sample to the sample holder by placing it on the adhesive coated side of a postit note that has been attached to the sample holder using only double sided tape. This process should be completed far away from sample head/tip. 34 35 5. Place sample holder onto sample stage and slide under scan head, avoiding contact with cantilever. Bumping the cantilever may damage the tip. Be sure tip is centered above sample. 36 37 38 39 5. Lower scan head by turning screws in small increments, ensuring that centering is maintained. This is mostly done by turning two screws simultaneously for one turn (counterclockwise), then turning the third screw one turn, repeating until tip is 1mm from sample surface. Keep an eye on tip distance while adjusting screws to avoid contact between tip and sample; this will likely break the tip. 40 Running the software 41 Starting the software: 42 43 44 45 Note: If the computer has never been used for AFM imaging, the software must first be installed. Insert the software CD and follow the Setup instructions. Also, note that the drivers for connection with the hardware will automatically be downloaded the first time the AFM will be switched on when connecting the PC. 46 1. Run the software by selecting start, all programs, Easyscan2, Nanosurf Easyscan 47 48 Should any error messages be displayed at this stage, look into the trouble shooting section in the Easyscan 2 manual. 49 50 51 52 2. Upon successful start of the program, click on “Positioning” in the leftmost part of the screen. A real time image should be displayed on the right side of the screen. Visual confirmation of the fact that the tip correctly mounted and not damaged is given by the presence of a needle-like structure projecting from the tip holder. If this structure is not present, the tip is almost certainly broken. 53 Tip Positioning: 54 55 56 57 1. Click the “Advance” button; this will cause the tip to lower onto the sample. You should see the tip’s shadow appear on the screen and move towards the tip. IMPORTANT NOTE: make sure to advance the tip in small increments; the tip MUST NOT touch the sample (i.e. the tip and its shadow should not touch during this operation). 58 Stop when the tip is separated from its shadow on the screen by about one centimeter. 59 60 2. Select the Mode of operation depending on the type of tip selected (which is also specified on the tip’s box). 61 For Sicion-A, select Static force 62 For ACL-A, select Tapping force 63 3. Click on Approach. The computer should automatically lower the tip completely onto the sample. 64 Imaging: 65 1. Click the “imaging” button on the leftmost part of the screen. 66 67 2. Before starting the scan (see next point), you can choose the initial values for Width (defaut:0.5), Time Line (default 1.0 seconds), and Points per line (default: 256) 68 3. Click on “Start” to begin scanning. 69 70 4. Adjust the P-gain and I-gain values in order to produce a smooth graph. Typical values are 5000 for the p-gain and 1000 for the I-gain. 71 5. Adjust the x-slope value until it gets close to zero. 72 6. Set the rotation value to 90 and repeat the previous step. 73 Shutting down Atomic Force Microscope 74 1. Stop scanning 75 76 2. Retract cantilever as far as possible by auto-positioning software feature. Leave software running in case needed for an unforeseen situation. 77 3. Remove sample from stage 78 4. Shut down software. 79 5. Turn off power to controller 80 81 6. Disconnect power cable, computer cable, scan head cable, video camera cable, and store in proper pockets. Place controller in proper pocket. 82 83 7. Remove scan head from stage and place in plastic case. Do not remove cantilever because of risk of damage is substantial. Only change cantilevers as needed. 84 8. Place scan head plastic case in proper pocket 85 86 9. Reinstall the four cardboard spacers in the corners of the anti-vibration table and tighten hex bolts approximately one quarter turn past tight. Do not over tighten. 87 10. Shut down PC and store AFM cases in proper shelves.