Klapperich Lab:Guide to Calibration

Overview

The following manual describes how to operate the Hysitron Triboindenter in quasistatic, dynamic, or imaging modes and acquire and analyze data. It also presents common pitfalls, troubleshooting techniques, and describes key differences in the testing of hard versus soft materials and uses of different tips.

Materials

• Appropriate Tip (Berkovich, Fluid Cell, etc...)
• Tweezers
• Superglue (if sample is not attached to AFM disk)
• Millipore Water (if using Fluid Cell Tip)

Procedure

Start Up

1. Reserve the TriboIndenter at Yahoo Calendars
   • User Name: nanoindenter
   • Password: hysitron
2. Turn on the computer if not already on

Changing the Tip

1. With the computer on and the TriboScan software closed, turn off the piezo and transducer controllers.
2. Unplug the left cable from the transducer holder and the granite horizontal beam .
3. Loosen the transducer holder by unscrewing the horizontal silver knob to the right of microscope.
4. Slide the transducer holdlder upwards and out of the hood.
5. Unscrew the tip using the tip tool and place in the hollow tube.
6. Screw the tip in you desire stopping at the first sign of resistance.
7. Slide the transducer holder back into place, connect the cables and retighten the screw.
8. Turn both the peizo and transducer controllers back on.
9. Rezero the tare value.
10. Start TriboScan software.
11. In the positioning window, go to Setup>>Tip Area Function. Select the correct tip area function.

Z-Axis Calibration

1. Change the microscope feedback gain to 100 on the TriboScope panel and in the software. From the Positioning window select Setup>>Piezo Controls and change the value for Front-panel Microscope Feedback Gain to 100. Minimize this window as a reminder to change the value back.
2. From the positioning window, go to Setup>>Indent Open Loop Load Function.
3. Set up the Load Function Editor to perform a 600uN 2-segment indent in air (AIR INDENT.ldf), for a total of 20 seconds. For soft materials, this load should be increased so that the air indent goes approximately 5um, but 600uN should be sufficient.
4. From the Open Loop Load Function window, select Setup>>Advanced z-axis Calibration.
5. Click "Calibrate Transducer." The instructions at this point from the pop-up windows are fairly self-explanatory.
6. The calibration window will pop up. Check that the following values are correct.

||Parameter||Correct Value|| ||Machine Compliance||4.0|| ||Displacement Gain||100|| ||Bias Offset||0.2|| ||Disp. & Micro. Feedback gain auto detect||Unchecked||

      ||row1-column1|row1-column2||row2-column1|row2-column2||

Notes

1. List troubleshooting tips here.
2. You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
3. Anecdotal observations that might be of use to others can also be posted here.

Please sign your name to your note by adding ('''~~~~''') to the end of your tip.

References

Relevant papers and books

1. Goldbeter A and Koshland DE Jr. An amplified sensitivity arising from covalent modification in biological systems. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6840-4. DOI:10.1073/pnas.78.11.6840 |
2. JACOB F and MONOD J. Genetic regulatory mechanisms in the synthesis of proteins. J Mol Biol. 1961 Jun;3:318-56. DOI:10.1016/s0022-2836(61)80072-7 |
3. ISBN:0879697164

Contact

• Who has experience with this protocol?