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3D imaging in CUBIC-cleared mouse heart tissue: going deeper.

Imke Nehrhoff¹, Diana Bocancea², Javier Vaquero³

¹Instituto de Investigación Sanitaria Gregorio Marañón. (IiSGM), Madrid, Spain; These authors contributed equally to this work.

Biomedical Optics Express

|October 5, 2016

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Researchers optimized the CUBIC clearing protocol for mouse hearts, improving 3D imaging depth and antibody penetration. This allows for more detailed studies of heart disease at the cellular level.

Area of Science:

Cardiovascular Biology
Biomedical Imaging
Histology

Background:

High-resolution 3D imaging is crucial for detailed study of heart diseases.
Existing methods face limitations in tissue penetration for thick samples.
Understanding cellular and structural changes requires advanced imaging techniques.

Purpose of the Study:

To optimize the CUBIC clearing protocol for thick mouse heart sections.
To enhance laser penetration depth for confocal microscopy.
To improve antibody penetration for detailed molecular analysis.

Main Methods:

Optimization of the CUBIC (clear, unobstructed brain imaging cocktails and computational analysis) clearing protocol.
Application of the protocol to thick mouse heart tissue sections.

Keywords:

(000.1430) Biology and medicine (180.1790) Confocal microscopy

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Last Updated: Mar 14, 2026

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Confocal microscopy for deep 3D image acquisition.

Assessment of antibody penetration efficiency.

Main Results:

Achieved enhanced penetration depth of confocal microscope lasers into thick mouse heart tissue.
Increased antibody penetration into the heart tissue by a factor of five.
Enabled high-quality, deep 3D image acquisition of the heart.

Conclusions:

The optimized CUBIC protocol significantly improves 3D imaging capabilities for mouse hearts.
This advancement facilitates more accurate assessment of cardiac cellular and structural changes.
The protocol is valuable for research into the mechanisms of heart diseases.