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The mounted samples were immersed in juice medium to obtain a constant potential, which is referred as the Open Circuit Potential.
The mounted samples were immediately transferred into the AFM liquid cell, while avoiding dewetting.
The mounted samples were imaged using anLSM 510 confocal microscope (Zeiss;Hamburg, Germany), with a 100× or a 40× objective lens.
The test began when the mounted samples were added to the saliva medium.
The mounted samples were then cleared through a series of 25%.
The mounted samples were stained with 1% toluidine blue for light microscopy.
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The fluorescence signal from the mounted samples was visualised under a fluorescence microscope with a × 20 objective.
After filtering a concentrated cell suspension, the filter was gently rinsed with buffer, carefully cut (1 cm ×1 cm), attached to a steel sample puck (Veeco Metrology Group) and the mounted sample was transferred into the AFM liquid cell while avoiding dewetting.
A scheme of the mounted sample is presented in Figure 1.
Whole-mount samples were documented with a Nikon SMZ1500 dissection microscope fitted with a Leica DC500 camera.
Whole-mount samples were then cryosectioned, washed in PBS and mounted in glycerol for imaging.
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