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Related Experiment Video

Updated: Jun 1, 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

Single-shot optical sectioning using two-color probes in HiLo fluorescence microscopy.

Eleonora Muro¹, Pierre Vermeulen, Andriani Ioannou

¹Laboratoire de Physique et d'Étude des Matériaux, Centre National de la Recherche Scientifique, UMR 8213, École Supérieure de Physique et de Chimie Industrielles, Paris, France.

Biophysical Journal

|June 7, 2011

Summary

This study introduces a novel wide-field fluorescence microscope combining HiLo microscopy and dual-color probes for rapid, one-shot optical sectioning of moving biological samples like Xenopus laevis embryos.

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Last Updated: Jun 1, 2026

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Area of Science:

Biophotonics
Microscopy
Developmental Biology

Background:

Traditional fluorescence microscopy struggles with thick, moving samples.
Optical sectioning is crucial for high-resolution imaging of biological structures.
Simultaneous multi-wavelength illumination is challenging for dynamic specimens.

Purpose of the Study:

To develop a wide-field fluorescence microscope for fast, optical sectioning of thick, moving biological samples.
To integrate HiLo microscopy with a two-color fluorescent probe for enhanced imaging.
To achieve one-shot imaging at high frame rates.

Main Methods:

A novel wide-field fluorescence microscope setup was designed.
The setup combines HiLo microscopy with simultaneous dual-wavelength illumination (flat and structured patterns).
Spectrally separated images were acquired and combined using the HiLo technique.

Main Results:

The system enables one-shot fluorescence optical sectioning of thick, moving samples.
Optically sectioned, full-field images of Xenopus laevis embryos were acquired.
High-speed imaging at a 25 frames/s rate was achieved.

Conclusions:

The developed microscope effectively performs optical sectioning on dynamic biological samples.
This technique offers a significant advancement for imaging thick, moving specimens in real-time.
The method is suitable for studying developmental processes in organisms like Xenopus laevis embryos.