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Fixation and Sectioning

Fixation 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...

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Related Experiment Video

Updated: Oct 15, 2025

Staining and High-Resolution Imaging of Three-Dimensional Organoid and Spheroid Models

Staining and High-Resolution Imaging of Three-Dimensional Organoid and Spheroid Models

Published on: March 27, 2021

Tutorial: methods for three-dimensional visualization of archival tissue material.

Tariq Sami Haddad¹, Peter Friedl^2,3,4, Navid Farahani⁵

¹Department of Pathology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, the Netherlands. tariq.haddad@radboudumc.nl.

Nature Protocols

|October 30, 2021

Summary

This summary is machine-generated.

This tutorial explores 3D visualization of archival tissue, offering methods to unlock valuable insights into tissue structure and disease mechanisms from historical samples.

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Area of Science:

Biomedical Imaging
Histopathology
3D Tissue Analysis

Background:

Three-dimensional (3D) analysis of patient specimens is crucial for understanding tissue structure and disease mechanisms.
Advancements in imaging technologies enable visualization of large tissue volumes at subcellular resolution.
Archival tissue samples, preserved for decades, represent an underutilized resource for scientific research.

Purpose of the Study:

To provide a comprehensive tutorial on sample preparation and microscopy techniques for 3D visualization of archival tissue.
To address common challenges associated with archival tissue analysis, such as registration and antibody penetration.
To guide researchers in processing, visualizing, and analyzing 3D imaging data from archival samples.

Main Methods:

Overview of sample preparation techniques tailored for archival tissues.
Discussion of various microscopy approaches suitable for 3D tissue visualization.
Guidance on data analysis pipelines, including registration and visualization strategies.

Main Results:

Key aspects of relevant techniques for 3D archival tissue visualization are summarized.
Common issues encountered, including antibody penetration and data registration, are addressed.
Criteria for decision-making in data analysis are provided.

Conclusions:

The outlined methods offer a sustainable approach to validating 3D tissue organization.
This tutorial facilitates the study of disease mechanisms using valuable archival tissue resources.
Enhanced 3D analysis of archival tissues can significantly advance biomedical research.