Sign In

Histological Sample Preparation for Light Microscopy

Overview

Histology is the study of cells and tissues, which is typically aided by the use of a light microscope. The preparation of histological samples can vary greatly based on the inherent properties of the samples such as size and hardness as well as expected post-processing which includes planned staining techniques or other down-stream applications. As described in this video, specimen preparation typically begins with a fixation procedure to prevent degradation of the sample by naturally occurring enzymes that are released by the cells upon death. Once fixed, samples are placed into an embedding medium that is able to sufficiently support the sample. Most commonly this is paraffin wax, but other materials such as a glycerin based freezing medium and agars are also used to surround the sample during sectioning. Sectioning then takes place on a microtome or other cutting device that allows the user to shave the sample into thin slices ranging from a few microns to a few millimeters in thickness. Once cut, sections are mounted on a glass slide and stained to bring out specific features of the sample before being imaged on a microscope.

Procedure

Histology is a term that refers to the study of the microscopic anatomy of tissues and cells. Proper histological sample preparation for light microscopy is essential for obtaining quality results from tissue samples.

There are 3 main steps common to nearly all histological procedures. First, the sample is fixed, in order to preserve the tissue and slow down tissue degradation. Next, the sample is immersed in a material, or embedding media, that has similar mechanical properties to itself. Once embedded, the sample

Log in or to access full content. Learn more about your institution’s access to JoVE content here

Tags
Histological Sample PreparationLight MicroscopyHistologyTissue SamplesFixingEmbedding MediaMicrotomeSectioningTissue FixationCell DegradationEnzymesProtein DegradationCross linkingCoagulationProtein Dehydration

Skip to...

0:00

Overview

1:05

Fixation of Tissue Samples

4:19

Embedding and Sectioning

6:12

Applications

9:01

Summary

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved