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Related Concept Videos

Fixation and Sectioning01:03

Fixation and Sectioning

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Two basic types of preparation are used to visualize specimens with a light microscope: wet mounts and fixed specimens.
The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...
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Related Experiment Video

Updated: Sep 24, 2025

Free-floating Immunostaining of Mouse Brains
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Histological-Based Stainings using Free-Floating Tissue Sections.

Emily M Potts1, Giuseppe Coppotelli1, Jaime M Ross2

  • 1George & Anne Ryan Institute for Neuroscience, College of Pharmacy, Department of Biomedical and Pharmaceutical Sciences, University of Rhode Island.

Journal of Visualized Experiments : Jove
|September 14, 2020
PubMed
Summary
This summary is machine-generated.

The free-floating method offers superior antibody penetration for immunohistochemistry, especially for thick brain sections used in 3D reconstruction. This technique simplifies handling, making it ideal for large-scale studies and various staining methods.

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Area of Science:

  • Histology
  • Neuroscience
  • Biomedical Research

Background:

  • Immunohistochemistry visualizes tissue structures and protein expression.
  • Two main methods exist: slide-mounted and free-floating sections.
  • Free-floating allows sections to be stained while suspended in solution.

Purpose of the Study:

  • To detail the application of the free-floating method for fluorescent immunohistochemistry.
  • To highlight its advantages for brain section analysis and 3D reconstruction.
  • To demonstrate its adaptability for other tissues and staining techniques.

Main Methods:

  • Utilizing the free-floating technique for tissue section staining.
  • Focusing on fluorescent immunohistochemistry of brain sections.
  • Adapting the method for various tissues and histochemical stains (H&E, cresyl violet).

Main Results:

  • The free-floating method enhances antibody penetration, crucial for thick sections.
  • It facilitates easier handling of multiple sections (30-40 per aliquot).
  • The technique is versatile and adaptable for diverse research needs.

Conclusions:

  • The free-floating technique is recommended for immunohistochemistry, particularly for thick sections and 3D reconstruction.
  • It offers practical advantages in handling and scalability for large-scale studies.
  • This method is adaptable to various tissues and histochemical staining protocols.