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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: Jun 4, 2026

Fluorescence Imaging with One-nanometer Accuracy (FIONA)

Fluorescence Imaging with One-nanometer Accuracy (FIONA)

Published on: September 26, 2014

High accuracy FIONA-AFM hybrid imaging.

D N Fronczek¹, C Quammen, H Wang

¹Rudolf Virchow Center for Experimental Biomedicine, University of Würzburg, Josef Schneider Strasse 2, 97080 Würzburg, Germany.

Ultramicroscopy

|February 19, 2011

Summary

This summary is machine-generated.

This study introduces FIONA-AFM, a novel technique combining fluorescence and atomic force microscopy to identify individual proteins within complexes. This method enhances structural analysis of multi-protein assemblies, advancing biological research.

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Fluorescence Imaging with One-nanometer Accuracy (FIONA)

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Published on: September 26, 2014

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Sub-nanometer Resolution Imaging with Amplitude-modulation Atomic Force Microscopy in Liquid

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Published on: December 20, 2016

Area of Science:

Biophysics
Molecular Biology
Nanotechnology

Background:

Multi-protein complexes are crucial for biological functions.
Current imaging methods like electron microscopy (EM) and atomic force microscopy (AFM) struggle to differentiate individual proteins within complexes.
Distinguishing proteins is vital for understanding complex structure and function.

Purpose of the Study:

To develop a hybrid imaging technique for identifying distinct proteins in multi-protein complexes.
To achieve high-accuracy spatial registration between fluorescence and AFM data.
To enable detailed structural and spatial analysis of protein interactions.

Main Methods:

Combining high-resolution fluorescence imaging with atomic force microscopy (AFM).
Utilizing quantum dots as fiducial markers for precise alignment.
Developing Fluorescence Imaging with One Nanometer Accuracy (FIONA)-AFM (FIONA-AFM).

Main Results:

Achieved alignment accuracy of ≥8nm between fluorescence and AFM data.
Demonstrated the ability to identify individual fluorescently labeled proteins within complexes.
Investigated the impact of localization precision and accuracy on hybrid image registration.

Conclusions:

FIONA-AFM successfully identifies individual proteins in multi-protein complexes.
The technique provides high-resolution structural insights (5-10nm) via AFM and spatial protein relationships via fluorescence.
This hybrid approach significantly expands the study of protein interactions and complex dynamics.