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The volume was measured in the regions between fiducial marks (yellow dotted outline).
The volume was measured in the regions between fiducial marks (red outline).
The final accuracy in retrieving track coordinates after fiducial marks measurement is better than 1μm.
The fiducial marks added an extra FN layer to the FN nanofibers and thus appear white in the AFM images.
For FN nanofibers that were uniaxially stretched, only the regions of the nanofibers between fiducial marks was masked and the volume was measured as previously described.
We first microcontact printed 1 cm long and 67 μm wide lines of FN that have been tagged with fiducial marks (Supplementary Figure 1).
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This fiducial mark was visible after resin embedding and was used to locate the imaged axon37,38.
To calculate strain, we measured the center-to-center fiducial mark distance of FN nanofibers in a pre-release state as the initial length and the center-to-center fiducial mark distance of strained FN nanofibers as the final length.
FN nanofibers pre-release on PIPAAm had a center-to-center fiducial mark distance of 20.21 ± 0.68 μm (Fig. 6b, n = 3).
With the feed-in position of the tool, the difference between the fiducial mark and the target position can be obtained by image processing.
When released, strained, and immobilized for AFM imaging, we measured an average fiducial mark center-to-center distance of 40.06 ± 8.68 μm (Fig. 6c, n = 5) representing an average strain of 98.2%.
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