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Updated: May 24, 2026

Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy (iPALM)

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Published on: December 1, 2016

PSF shaping using adaptive optics for three-dimensional single-molecule super-resolution imaging and tracking.

Ignacio Izeddin¹, Mohamed El Beheiry, Jordi Andilla

¹Laboratoire Kastler Brossel, CNRS UMR 8552, Département de Physique et Institut de Biologie de l’Ecole Normale Supérieure, Université Pierre et Marie Curie-Paris6, 46 rue d’Ulm 75230 Paris cedex 05, France.

|March 16, 2012

Summary

We developed a new adaptive optics method for precise 3D single-molecule localization. This technique achieves nanometer-level accuracy for fluorescent molecules, enabling high-resolution imaging and tracking in biological samples.

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Last Updated: May 24, 2026

Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy (iPALM)

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Area of Science:

Biophysics
Optical Microscopy
Nanotechnology

Background:

Accurate three-dimensional (3D) localization of single molecules is crucial for understanding cellular processes.
Conventional microscopy techniques often face limitations in achieving high axial resolution and precision.
Adaptive optics offers potential for overcoming optical aberrations and enhancing imaging performance.

Purpose of the Study:

To present a novel adaptive optics approach for precise 3D single-molecule localization.
To investigate the impact of induced astigmatism on z-localization precision.
To demonstrate the utility of this method in super-resolution microscopy and live-cell imaging.

Main Methods:

Utilizing a 52-actuator deformable mirror to correct aberrations and introduce controlled astigmatism in the point-spread-function.
Characterizing the relationship between astigmatism and z-localization precision.
Applying the technique to image fixed cells and track labeled proteins in live cells.

Main Results:

Achieved a z-localization precision of 40 nm for fluorescent proteins and 20 nm for fluorescent dyes.
Demonstrated precise localization over an axial depth of approximately 800 nm.
Generated super-resolution images of actin filaments and performed single-molecule tracking of transmembrane proteins.

Conclusions:

The developed adaptive optics method significantly enhances 3D single-molecule localization precision.
This technique is effective for both static super-resolution imaging and dynamic single-molecule tracking in live cells.
The approach holds promise for advancing high-resolution biological imaging and molecular studies.