Automated setpoint adjustment for biological contact mode atomic force microscopy imaging.
Contact mode atomic force microscopy (AFM) is the most frequently used AFM imaging mode in biology. It is about 5-10 times faster than oscillating mode imaging (in conventional AFM setups), and provides topographs of biological samples with sub-molecular resolution and at a high signal-to-noise ratio. Unfortunately, contact mode imaging is sensitive to the applied force and intrinsic force drift: inappropriate force applied by the AFM tip damages the soft biological samples. We present a methodology that automatically searches for and maintains high resolution imaging forces. We found that the vertical and lateral vibrations of the probe during scanning are valuable signals for the characterization of the actual applied force by the tip. This allows automated adjustment and correction of the setpoint force during an experiment. A system that permanently performs this methodology steered the AFM towards high resolution imaging forces and imaged purple membrane at molecular resolution and live cells at high signal-to-noise ratio for hours without an operator.
Casuso
Ignacio
I
Institut Curie, Equipe INSERM Avenir, UMR168-CNRS, Paris, France.
Scheuring
Simon
S
D001331:Automation; D001696:Biomechanical Phenomena; D019617:Halobacterium salinarum / Q000648:ultrastructure; D018625:Microscopy, Atomic Force / Q000379:methods*; D018717:Purple Membrane / Q000648:ultrastructure*; D055213:Retinal Pigment Epithelium / Q000166:cytology; D013997:Time Factors; D014732:Vibration