Universidad Politécnica de Sinaloa

Ing. En Nanotecnología

English Vlll

Teacher: Stefany Ramírez Parra

Velazquez Vázquez Humberto

Hinojosa Moran Pedro Jesús

Solís Vázquez Erick Gpe

Balderas Torres Roberto

23/01/2020
 The AFM
Atomic force microscopy is arguably the most versatile and powerful microscopy technology
for studying samples at nanoscale. It is versatile because an atomic force microscope can not
only image in three-dimensional topography, but it also provides various types of surface
measurements to the needs of scientists and engineers. It is powerful because an AFM can
generate images at atomic resolution with angstrom scale resolution height information, with
minimum sample preparation.

AFM Principle - Surface

Sensing: An AFM uses a
cantilever with a very sharp tip to
scan over a sample surface. As
the tip approaches the surface,
the close-range, attractive force
between the surface and the tip
Figure 1: AFM Components
cause the cantilever to deflect
towards the surface. However, as the cantilever is brought even closer to the surface, such
that the tip makes contact with it, increasingly repulsive force takes over and causes the
cantilever to deflect away from the surface.

- Detection Method: A laser beam is used to detect cantilever deflections towards or away
from the surface. By reflecting an incident beam off the flat top of the cantilever, any
cantilever deflection will cause slight changes in the direction of the reflected beam. A
position-sensitive photo diode (PSPD) can be used to track these changes. Thus, if an AFM
tip passes over a raised surface feature, the resulting cantilever deflection (and the subsequent
change in direction of reflected beam) is recorded by the PSPD.

- Imaging: An AFM images the topography of a sample surface by scanning the cantilever
over a region of interest. The raised and lowered features on the sample surface influence the
deflection of the cantilever, which is monitored by the PSPD. By using a feedback loop to
control the height of the tip above the surface thus maintaining constant laser position the
AFM can generate an accurate topographic map of the surface features.
 AFM Setup

1. Switch on the computer and the vibration isolation system, if not already on.

2. Turn the laser on if the key switch is in the OFF position.

3. Switch on the AFM controller.

4. Launch IGOR Pro.

5. Remove fluid chamber from the AFM.

6. Mount a cantilever in the fluid chamber’s clamp (Figure 2). Use plastic tweezers to avoid
scratching the glass. Make sure the cantilever is aligned with its holder (yellow rectangle in
Figure 2) and totally inside the receptacle. A dissecting microscope or magnifying glass can
be used for this purpose.

Figure 2: Cell System. H1 Fluid chamber; H2 Cell Slider; H3 Holder; H4 Insert; S3 Fixing knob for X
adjustment; S5 Screw for Y adjustment
7. Place the fluid chamber back on the AFM and secure it with the S9 screw (Figure 2).

8. Align the laser using the AFM optic system.

9. Load the tempered buffer onto the fluid chamber and check that the laser and the cantilever
tip are still aligned. Repeat step 8 if necessary, making sure that the buffer is covering the
cantilever completely.

10. If the coverslip from the previous experiment is still in place, remove it. Clean the surface
of the piezo with 70% ethanol using a wet piece of paper. Make sure the surface is dry before
proceeding to next step.

11. Coat the piezo with a thin layer of sterile grease and place a compressed air‐cleaned (gold,
chloroalkane, etc) coverslip on it. Even though the coverslip can be cleaned this way, NEVER
use compressed air on the piezo, since it may damage it. Always clean the coverslip BEFORE
putting it on the piezo.

12. After centrifuging the protein sample, apply the desired volume on top of the coverslip
and gently spread it to create a thin layer as done before with the grease. Use the pipette tip
for this, always trying not to scratch the surface.

13. Slowly turn the piezo upside down until it faces the fluid chamber.
Warnings
 Diagrams

Figure 3: AFM Components

Figure 4: How the laser beam works in a AFM