HISTOLOGICAL TECHNIQUES
DISCIPLINE OF PHYSIOLOGY
INTRODUCTION TO HISTOLOGY
ANAT203W2
September2021
Compiled by: D A Margolis
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Histological Techniques: Preserving Microscopic Anatomy, Cheat Sheet of Neurology
This document provides a comprehensive overview of histological techniques essential for preparing tissue samples for microscopic examination. It covers the key processes involved, including fixation, dehydration, clearing, impregnating, embedding, sectioning, and staining, to preserve the microscopic anatomy of the tissue as close to its natural state as possible. The document also explains the principles behind different types of dyes and their interaction with cellular components, enabling effective tissue preparation and microscopic analysis.
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HISTOLOGICAL TECHNIQUES
DISCIPLINE OF PHYSIOLOGY INTRODUCTION TO HISTOLOGY ANAT203W September Compiled by: D A Margolis
What is Histotechnique?
- Histological technique deals with the preparation of tissue for microscopic
examination.
- The aim of good histological technique to preserve microscopic anatomy
(histology) of tissue.
What are the Processes/Techniques?
- These techniques are:
- Fixation
- Dehydration
- Cleaning/Clearing
- Impregnating & Embedding
- Sectioning (Microtomy)
- Staining
Fixing Tissue
- Fixation to preserve tissue and maintain “life –like” structure.
- Take fresh or perfused tissue
- Cut into 1cm cube
- Place into fixative e.g. 10% formalin
- Leave to fix for 12hours
- After fixation, place tissue into marked tissue processing
cassettes
Blocking/Embedding
- Remove cassettes containing tissue from the tissue processor
- Fill wax mould with wax from the dispenser
- Remove tissue sample from cassette with warm forceps and place in
the mould.
- Place cassette base on top of mould and fill with wax.
- Place on a cold plate for 30 minutes.
- Remove the blocked sample from the mould.
Sectioning the Sample
- This produces sections thin enough to allow viewing through a
microscope.
- Use a rotary microtome to cut sections 3-5 microns in thickness.
- Float tissue sections in warm water to flatten
- Pick sections up onto a microscopic glass slide.
- Allow to dry at to ensure, sections adhere to the slide.
Acid Dyes
In an acid dye the basic component is colored and the acid component is
colorless.
Acid dyes stain basic components e.g. eosin stains cytoplasm.
The color imparted is shade of red.
Basic Dyes
- In a basic dye the acid component is colored and the basic component is
colorless.
- Basic dyes stain acidic components e.g. hematoxylin nucleus.
- The color imparted is shade of blue.