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Updated: Jun 26, 2026

Fluorescence Imaging with One-nanometer Accuracy (FIONA)

Fluorescence Imaging with One-nanometer Accuracy (FIONA)

Published on: September 26, 2014

Chapter 12: Nanoscale biological fluorescence imaging: breaking the diffraction barrier.

Travis J Gould¹, Samuel T Hess

¹Department of Physics and Astronomy and Institute for Molecular Biophysics, University of Maine, Orono, Maine 04469, USA.

Methods in Cell Biology

|January 3, 2009

Summary

Ultra-high-resolution microscopy, such as Fluorescence Photoactivation Localization Microscopy (FPALM), overcomes light diffraction limits. This technique achieves nanoscale resolution for imaging cellular structures in biological samples.

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Fluorescence Imaging with One-nanometer Accuracy (FIONA)

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Area of Science:

Cell Biology
Microscopy
Biophysics

Background:

Traditional light microscopy is constrained by the diffraction limit, restricting resolution in biological imaging.
Advancements in fluorescence-based techniques are enabling super-resolution imaging, surpassing conventional limitations.
Imaging intracellular protein distributions at the nanoscale is crucial for understanding cellular functions.

Purpose of the Study:

To present a detailed description of Fluorescence Photoactivation Localization Microscopy (FPALM) methods for biological samples.
To demonstrate the capability of FPALM for ultra-high-resolution imaging.
To compare FPALM with other existing super-resolution microscopy techniques.

Main Methods:

Utilizes fluorescence photoactivation and localization principles to achieve super-resolution.
Applies the technique to image both living and fixed biological cells.
Detailed methodology for sample preparation and image acquisition is provided.

Main Results:

Achieved lateral resolution of better than 20 nanometers.
Demonstrated the ability to visualize fine intracellular protein distributions.
FPALM effectively breaks the diffraction barrier in light microscopy.

Conclusions:

FPALM is a powerful ultra-high-resolution microscopy method for biological imaging.
The technique offers significant improvements over conventional light microscopy.
FPALM provides new possibilities for studying cellular ultrastructure at the nanoscale.