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Updated: Jul 13, 2026

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

Parallel two-channel near- and far-field fluorescence microscopy.

Dorinel Verdes¹, Thomas Ruckstuhl, Stefan Seeger

¹Universität Zürich, Physikalisch-Chemisches Institut, Winterthurerstrasse 190, Zürich CH-8057, Switzerland.

Journal of Biomedical Optics

|July 7, 2007

Summary

This study introduces a novel two-channel fluorescence microscopy method for high-resolution imaging of aqueous samples. The technique simultaneously captures surface-bound and unbound molecules using distinct detection volumes.

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Conducting Multiple Imaging Modes with One Fluorescence Microscope

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Published on: October 28, 2018

Area of Science:

Biophysics
Optical Microscopy
Surface Science

Background:

Accurate imaging of molecular interactions at interfaces is crucial.
Conventional microscopy techniques struggle to differentiate surface-bound from unbound molecules in aqueous environments.

Purpose of the Study:

To develop a novel fluorescence microscopy technique for high-resolution imaging of aqueous samples.
To enable simultaneous detection of surface-bound and unbound fluorescent molecules.

Main Methods:

A two-channel fluorescence microscope utilizing a parabolic mirror objective and an aspheric lens was developed.
Supercritical angle fluorescence (SAF) and subcritical angle fluorescence were collected separately.
Detection volumes with differing axial resolutions were generated at the glass-water interface.

Main Results:

The technique allows for high-resolution imaging of aqueous samples.
Supercritical angle fluorescence collection achieved strict surface confinement of the detection volume.
Subcritical angle collection enabled detection of fluorescence several microns deep within the sample.

Conclusions:

The developed fluorescence microscopy technique successfully distinguishes between surface-bound and unbound fluorescent molecules.
This method provides simultaneous acquisition of signals from different depths, enhancing surface analysis capabilities.