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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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Related Experiment Video

Updated: Jun 13, 2026

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

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"Seeing and counting" individual antigens captured on a microarrayed spot with force-based atomic force microscopy.

Dhruvajyoti Roy¹, Sung Hong Kwon, Ju-Won Kwak

¹Department of Chemistry, National Core Research Center for Systems Bio-Dynamics, Pohang University of Science and Technology (POSTECH), Pohang 790-784, Korea.

Analytical Chemistry

|May 18, 2010

Summary

This summary is machine-generated.

Atomic force microscopy (AFM) precisely quantifies prostate-specific antigens (PSAs) on microarrays. This method enhances reliability for detecting low PSA concentrations, crucial for early prostate cancer detection.

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Counting Proteins in Single Cells with Addressable Droplet Microarrays

Counting Proteins in Single Cells with Addressable Droplet Microarrays

Published on: July 6, 2018

Area of Science:

Nanotechnology
Biophysics
Analytical Chemistry

Background:

Accurate quantification of biomarkers like prostate-specific antigens (PSAs) is vital for disease diagnosis.
Existing methods face challenges in sensitivity and reproducibility for low analyte concentrations.

Purpose of the Study:

To develop and validate a highly sensitive and reproducible atomic force microscopy (AFM)-based method for quantifying prostate-specific antigens (PSAs).
To assess the impact of surface modification using dendrons on the reliability of PSA detection.

Main Methods:

Utilized atomic force microscopy (AFM) for high-resolution imaging and force measurements of captured prostate-specific antigens (PSAs) on antibody-microarrayed spots.
Employed a third-generation dendron for surface treatment to enhance measurement reliability and reproducibility.
Measured the specific unbinding force between captured PSA and the detection antibody (5A6).
Analyzed the number of captured antigens within a submicrometer area at varying concentrations.

Main Results:

The mean unbinding force between captured PSA and the detection antibody was determined to be 56 +/- 2 pN.
At 100 fM, 12 captured antigens were detected in a 4.32 x 10(4) nm(2) area, demonstrating concentration-dependent quantification.
A larger hydrodynamic distance (8 +/- 2 nm) of the immunocomplex facilitated discrimination between specific and nonspecific interactions through pixel clustering.

Conclusions:

The developed AFM approach offers a reliable method for the quantitative analysis of antigens in biological samples.
The technique shows promise for detecting very low copy numbers of analytes, particularly when combined with reduced microarray spot sizes.
Surface functionalization with dendrons significantly improves the robustness of the nanoscale immunoassay.