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Related Experiment Video

Updated: Jul 29, 2025

Histological-Based Stainings Using Free-Floating Tissue Sections

Histological-Based Stainings Using Free-Floating Tissue Sections

Published on: August 25, 2020

A novel immunohistochemical protocol for paraffin embedded tissue sections using free-floating techniques.

Carolina Muniz Partida¹, Eric Walters¹

¹Department of Biochemistry and Molecular Biology, Howard University College of Medicine, Washington, DC, United States.

Frontiers in Neuroanatomy

|May 26, 2023

Summary

We developed a novel free-float immunohistochemistry protocol for paraffin-embedded tissues (PFFP). This method enhances tissue analysis, saving time, resources, and tissue samples.

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Area of Science:

Neuroscience
Histology
Biochemistry

Background:

Immunohistochemistry (IHC) is crucial for visualizing tissue and protein expression.
Current free-floating IHC methods use fragile cryo- or vibratome sections with poor morphology.
Limited information exists on free-floating IHC for paraffin-embedded tissues.

Purpose of the Study:

To develop and validate a free-float immunohistochemistry protocol for paraffin-embedded tissue (PFFP).
To assess PFFP's efficiency in localizing specific protein markers in various mouse brain regions.
To demonstrate PFFP's compatibility with different detection methods and downstream applications.

Main Methods:

Developed a novel free-float protocol for paraffin-embedded tissue (PFFP).

Keywords:

immunodetection immunofluorescence mouse brain neuroscience neurotransmitters

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Published on: October 7, 2021

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Last Updated: Jul 29, 2025

Histological-Based Stainings Using Free-Floating Tissue Sections

Histological-Based Stainings Using Free-Floating Tissue Sections

Published on: August 25, 2020

The Cutting and Floating Method for Paraffin-embedded Tissue for Sectioning

The Cutting and Floating Method for Paraffin-embedded Tissue for Sectioning

Published on: September 5, 2018

Free-floating Immunostaining of Mouse Brains

Free-floating Immunostaining of Mouse Brains

Published on: October 7, 2021

Applied PFFP to mouse brain sections (hippocampus, olfactory bulb, striatum, cortex).

Utilized chromogenic DAB and immunofluorescence detection with and without antigen retrieval.

Main Results:

PFFP successfully localized GFAP, olfactory marker protein, tyrosine hydroxylase, and Nestin.
Effective antigen localization was achieved with and without antigen retrieval.
Demonstrated PFFP's adaptability for various detection methods.

Conclusions:

PFFP offers a time-, resource-, and tissue-saving alternative to traditional IHC methods.
The PFFP protocol expands the utility of paraffin-embedded tissues for advanced analyses.
PFFP is compatible with in situ hybridization, protein interaction studies, and pathological diagnosis.