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Updated: May 28, 2026
07:41
Rigid Embedding of Fixed and Stained, Whole, Millimeter-Scale Specimens for Section-free 3D Histology by Micro-Computed Tomography
Published on: October 17, 2018
Fixation, embedding, and sectioning of tissues, embryos, and single cells
1Utrecht University, Utrecht, The Netherlands.
Current Protocols in Pharmacology
|October 4, 2011
Summary
This guide details methods for fixing and sectioning biological samples, including tissues and cells. These techniques prepare specimens for morphological examination and advanced analyses like immunohistochemistry.
Area of Science:
- Biological Sciences
- Histology
- Cell Biology
Background:
- Accurate preparation of biological samples is crucial for downstream analyses.
- Standardized protocols are needed for consistent tissue and cell sectioning.
- Various techniques exist for fixation and embedding, each with specific applications.
Purpose of the Study:
- To describe selected methods for fixing and sectioning diverse biological materials.
- To provide protocols for preparing sections suitable for morphological and molecular studies.
- To outline procedures for both tissue and cell-based sample preparation.
Main Methods:
- Fixation of tissues and embryos using paraformaldehyde followed by wax embedding.
- Alternative protocol for fixation of suspended or cultured cells.
- Perfusion of whole animals with paraformaldehyde for organ preservation.
- Sectioning of wax-embedded tissues and mounting onto glass slides.
Main Results:
- Established protocols for preparing fixed biological sections.
- Demonstrated applicability of sections for examining cell and tissue morphology.
- Validated use of sections in in situ hybridization, immunohistochemistry, and enzyme histochemistry.
- Provided methods for organ preservation and slide preparation.
Conclusions:
- The described methods offer a comprehensive approach to biological sample fixation and sectioning.
- Prepared sections are suitable for a range of histological and molecular investigations.
- These protocols support reproducible research in cell biology and tissue analysis.
Related Concept Videos
01:03Fixation and Sectioning
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...
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...
01:20Preparation of Samples for Electron Microscopy
To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...