45. Go to ‘File’ menu in the main toolbar and select ‘Live Scan’.
    
    
46. Click the ‘Approach’ button in the ‘Scan and Approach Control’ window. When the instrument reaches the set point, a dialogue box prompting ‘Setpoint Reached: Servo Active’ is displayed.
    
    
47. Click OK and go to the Scan tab and set the following:
    
    

    a. Scan size (area).

    b. Scan speed (number of lines/sec)

    c. Direction of scan (Up/Down/Toggle)

    d. Number of scans (Single/Multiple)

 

48. Go to the ‘Advanced Scan' tab and set the ‘Datapoints per line' (dpi).
    
    
49. Click Start to begin the scan (Note 1).
    
    
50. Save the obtained image as diphenylalanine.STP file (Note 2).
    
    
51. Go to the ‘Withdraw’ tab and click on ‘Withdraw’ to withdraw the probe.
    
    
52. Exit the ‘PicoScan’ software.
    
    
53. Remove the sample stage and unmount the mica sheet from the metallic disc.
    
    
54. Switch off the instrument as per manufacturer’s instructions.

Results and analysis:

1. Start the ‘PicoScan’ software.
   
   
2. Open the recorded image, the diphenylalanine.STP file.
   
   
3. In the ‘Data rendering’ window, go to the 3D scale tab and adjust the Z-scale to see the sample features.
   
   
4. In the same window (Data rendering), go to the ‘Optimizing' tab and flatten the image to obtain a flat background. This results in a clearer image on a flatter background.
   
   
5. A number of data processing options are present in the software. It is highly recommended to go through the software manual to realize its full potential.

Notes:

1. While the imaging is underway, any one of the image modes like Raw, Derivative, Flattened, and Tilted (in the ‘Optimizing' tab under ‘Data rendering' window) can be selected to obtain the image as per requirement. It is, however, recommended to obtain a raw image as any other processing can be done on the recorded image.
   
   
2. If the imaging is not good or the desired features are not obtained, it is recommended to scan a different area on the sample. This can be done without withdrawing the probe and giving an offset for the scanning area.