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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: Mar 16, 2026

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

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Visualization and Quantification of MicroRNA in a Single Cell Using Atomic Force Microscopy.

Hyunseo Koo¹, Ikbum Park¹, Yoonhee Lee¹

¹Department of Chemistry, ‡Division of Integrative Biosciences and Biotechnology, and §Department of Life Sciences, Pohang University of Science and Technology , 77 Cheongam-Ro, Nam-Gu, Pohang 37673, Korea.

Journal of the American Chemical Society

|August 17, 2016

Summary

We developed a sensitive atomic force microscopy (AFM) method to quantify microRNAs (miRNAs) in single cells. This technique accurately maps miRNA distribution and expression levels, advancing our understanding of cellular processes and diseases like cancer.

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Published on: July 10, 2019

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Area of Science:

Molecular Biology
Nanotechnology
Cell Biology

Background:

MicroRNAs (miRNAs) are crucial regulators of cellular processes.
Altered miRNA expression is linked to pathological conditions, including cancer.
Accurate single-cell miRNA quantification is essential for understanding miRNA function.

Purpose of the Study:

To develop a direct and sensitive method for single-cell miRNA detection.
To quantify miRNA copy number and map their distribution at the nanoscale.
To investigate changes in miRNA expression upon cellular activation.

Main Methods:

Utilized atomic force microscopy (AFM) with a hybrid binding domain (HBD)-tethered tip.
Developed an adhesion force mapping technique for individual miRNA localization.
Quantified miR-134 copy number in single neurons and fixed neuronal samples.

Main Results:

Successfully located and quantified mature miRNAs, distinguishing them from premature forms.
Demonstrated increased miR-134 expression in activated neurons (8-14 copies per 1.0 × 1.0 μm²).
Achieved nanometric lateral resolution mapping of miRNA distribution without amplification.

Conclusions:

AFM provides a reliable method for single-cell miRNA quantification and nanoscale mapping.
The technique enables the study of miRNA dynamics in response to cellular stimuli.
Sequential scanning with different AFM tips allows for multi-analyte analysis (miRNA, mRNA, proteins) for comprehensive post-transcriptional regulation studies.