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Updated: Jun 14, 2025

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

Simultaneous multicolor fluorescence imaging using PSF splitting.

Robin Van den Eynde¹, Fabian Hertel^1,2, Sergey Abakumov¹

¹Department of Chemistry, KU Leuven, Leuven, Belgium.

|September 6, 2024

Summary

This summary is machine-generated.

We developed a new fluorescence imaging method by splitting the point spread function (PSF). This technique enables simultaneous multicolor super-resolution and single-molecule microscopy across the entire field of view.

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Last Updated: Jun 14, 2025

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Published on: September 4, 2013

Area of Science:

Optical microscopy
Biophysics
Super-resolution imaging

Background:

Fluorescence imaging is crucial for biological research.
Current methods face limitations in multiplexing and field of view.
Point Spread Function (PSF) engineering offers potential for enhanced imaging.

Purpose of the Study:

To develop a novel method for encoding more information in fluorescence imaging.
To enable simultaneous multicolor super-resolution and single-molecule microscopy.
To maintain compatibility with existing imaging techniques and analysis tools.

Main Methods:

Splitting the original point spread function (PSF) to encode information.
Utilizing an add-on device called the 'Circulator'.
Encoding the fluorophore emission band into the PSF.

Main Results:

Demonstrated a method for broadband operation in fluorescence imaging.
Achieved simultaneous multicolor super-resolution and single-molecule microscopy.
Maintained compatibility with existing PSF engineering modalities and analysis tools.
Enabled imaging across essentially the full field of view.

Conclusions:

The developed method enhances information capacity in fluorescence imaging.
The 'Circulator' add-on facilitates advanced multicolor super-resolution microscopy.
This approach broadens the applicability of PSF engineering for biological imaging.