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Conducting Multiple Imaging Modes with One Fluorescence Microscope

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Published on: October 28, 2018

Isotropic 3D Nanoscopy based on single emitter switching.

Claas von Middendorff¹, Alexander Egner, Claudia Geisler

¹Max-Planck-Institute, Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany.

|December 10, 2008

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This study introduces a novel far-field fluorescence nanoscopy method achieving 5-10nm isotropic resolution by localizing individual emitters. This technique significantly enhances 3D imaging capabilities for advanced nanoscale visualization.

Area of Science:

Optical Microscopy
Nanotechnology
Biophysics

Background:

Far-field fluorescence nanoscopy faces challenges in achieving isotropic 3D resolution.
Existing methods often struggle with resolution limits in all spatial dimensions.

Purpose of the Study:

To develop and analyze a method for isotropic super-resolution in far-field fluorescence nanoscopy.
To achieve 3D resolutions down to 5-10nm in all directions.

Main Methods:

Employing emitter switching and mathematical localization of individual molecules.
Utilizing coherent detection of fluorescence light through opposing high-angle lenses.
Analyzing resolution using the Fisher information matrix.

Main Results:

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Demonstrated isotropic 3D resolution of 5-10nm under typical imaging conditions.
Provided a detailed theoretical analysis of resolution limits for single-molecule switching methods.
Verified results through Monte Carlo simulations and maximum-likelihood estimation.

Conclusions:

The proposed method effectively achieves isotropic super-resolution in far-field fluorescence nanoscopy.
This approach offers significant improvements for 3D nanoscale imaging.
The findings provide a robust framework for understanding and improving nanoscopy resolution.