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Related Experiment Video

Updated: Feb 6, 2026

Technique to Target Microinjection to the Developing Xenopus Kidney

Technique to Target Microinjection to the Developing Xenopus Kidney

Published on: May 3, 2016

X-FaCT: Xenopus-Fast Clearing Technique.

Pierre Affaticati¹, Sébastien Le Mével², Arnim Jenett¹

¹Tefor Core Facility, Paris-Saclay Institute of Neuroscience, CNRS, Université Paris-Saclay, Gif-sur-Yvette, France.

Methods in Molecular Biology (Clifton, N.J.)

|August 29, 2018

Summary

We developed a simplified fructose-based high-refractive index (fbHRI) solution for rapid 3D tissue clearing and immunolabeling of small biological specimens, reducing experimental time and tissue deformation.

Keywords:

3D imaging Fructose-based high-refractive index solution Tissue clearing

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Area of Science:

Cellular biology
Microscopy
Biochemistry

Background:

3D imaging of cellular structures in large specimens is essential for biological research.
Existing methods like passive CLARITY technique (PACT) enable in-toto immunolabeling and clearing of organs.
Previous work demonstrated tissue clearing on tadpole brains.

Purpose of the Study:

To present a simplified protocol for clearing small tissue samples (under 500 μm).
To reduce overall procedure time and minimize tissue deformation.
To adapt the protocol for thick tissue sections.

Main Methods:

Immersion of small tissue samples in a fructose-based high-refractive index (fbHRI) solution.
Refinement of protocol steps to accelerate the process.
Application of immunohistochemistry using anti-pH 3 antibody on Xenopus laevis brains.

Main Results:

Reduced overall procedure time by two thirds.
Enabled tissue clearing and immunolabeling within a week.
Minimized procedure-induced tissue deformations.
Successfully stained chromatin condensation associated with proliferation using anti-pH 3 antibody.

Conclusions:

The simplified fbHRI protocol offers a faster and more robust method for 3D imaging of small biological specimens.
This technique is easily adaptable for thick tissue sections.
The method facilitates detailed cellular analysis and reduces experimental burden.