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Atomic Force Microscopy

Atomic Force Microscopy

Atomic force microscopy (AFM) is a type of scanning probe microscopy that can analyze topographic details of various specimens like ceramics, glass, polymers, and biological samples. AFM offers over 1000 times more resolution than the optical imaging system. Images generated from AFM are three-dimensional surface profiles, offering an advantage over the flat, two-dimensional images from other imaging techniques.
The AFM Probe
The probe is regarded as the heart of any AFM setup and comprises the...

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Updated: May 1, 2026

Force Spectroscopy of Single Protein Molecules Using an Atomic Force Microscope

Force Spectroscopy of Single Protein Molecules Using an Atomic Force Microscope

Published on: February 28, 2019

Improved single molecule force spectroscopy using micromachined cantilevers.

Matthew S Bull¹, Ruby May A Sullan, Hongbin Li

¹JILA, National Institute of Standards and Technology and University of Colorado , Boulder, Colorado 80309, United States.

|March 28, 2014

Summary

This summary is machine-generated.

Researchers improved atomic force microscopy (AFM) force measurements by modifying cantilevers. This enhances precision and stability for studying biomolecules like DNA and proteins with single molecule force spectroscopy (SMFS).

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Force Spectroscopy of Single Protein Molecules Using an Atomic Force Microscope

Force Spectroscopy of Single Protein Molecules Using an Atomic Force Microscope

Published on: February 28, 2019

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Functionalization of Atomic Force Microscope Cantilevers with Single-T Cells or Single-Particle for Immunological Single-Cell Force Spectroscopy

Published on: July 10, 2019

Area of Science:

Atomic Force Microscopy (AFM)
Single Molecule Force Spectroscopy (SMFS)
Nanoscale Biophysics

Background:

Atomic force microscopy (AFM) is crucial for probing biomolecular dynamics and energetics, with single molecule force spectroscopy (SMFS) sensitive to sub-piconewton (pN) force changes.
Previous methods achieved sub-pN stability but compromised short-term force noise, temporal response, and sensitivity.
Gold-coated cantilevers offer reflectivity but can induce long-term drift, hindering precise measurements.

Purpose of the Study:

To enhance short-term force precision and long-term force stability in AFM for improved SMFS performance.
To overcome the trade-offs associated with previous cantilever modifications for AFM.
To develop robust and reusable cantilevers for broad AFM applications.

Main Methods:

Modification of short (40 μm) cantilevers using a focused ion beam.
Reduction of cantilever stiffness and hydrodynamic drag near a surface.
Preservation of cantilever reflectivity while mitigating gold-coating induced drift.

Main Results:

Achieved a ~10-fold reduction in cantilever stiffness and hydrodynamic drag.
Extended the sub-pN force bandwidth by approximately 50-fold (0.01–1000 Hz).
Demonstrated improved force precision and state-of-the-art stability in protein stretching experiments without loss of temporal resolution.

Conclusions:

Focused ion beam modification of short cantilevers offers superior AFM force measurement capabilities.
These modified cantilevers provide enhanced precision and stability for SMFS and other AFM modalities.
The cantilevers are robust, reusable, and expected to benefit diverse AFM-based research.