Fixation of special substances
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Formaldehyde and glutaraldehyde are commonly used crosslinking fixatives that terminate biochemical reactions and preserve tissues from decay by creating covalent bonds between proteins. Different fixatives are chosen based on the target molecule - for example, frozen sections are used to preserve lipids, while alcoholic fixatives are used for glycogen fixation. The choice of fixative also depends on subsequent testing methods.
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This document discusses tissue fixation in histopathology. It describes the various stages of histopathology including fixation, processing, embedding, sectioning and staining of tissues. It explains the importance of fixation in preventing autolysis and putrefaction of tissues. The key properties and reactions of common fixatives like formaldehyde and glutaraldehyde are outlined. Factors that can affect fixation quality including temperature, pH, concentration and duration of fixation are also summarized. Finally, different classifications of fixatives are presented based on their structural and functional properties.
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Fixatives and fixation/certified fixed orthodontic courses by Indian dental a...
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2) Clearing with xylene or other agents to make tissues transparent before wax infiltration.
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This document outlines the histotechnique process which tissues undergo before microscopic examination. Key steps include: fixation to preserve tissue structure; processing involving dehydration, clearing, and impregnation to allow sectioning; embedding tissues in paraffin blocks for microtomy; sectioning samples and staining, typically with hematoxylin and eosin, for visualization under the microscope. Finally, samples are mounted on slides and labeled for storage and pathological examination.
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The document summarizes the process of tissue processing which involves fixing, dehydrating, clearing, infiltrating with wax, embedding, sectioning, staining, and mounting tissue samples in order to examine them microscopically. Key steps include fixation to prevent decay, dehydration using alcohol to remove water, clearing with xylene to remove alcohol, infiltration and embedding in paraffin wax, sectioning thin slices with a microtome, staining typically with hematoxylin and eosin for examination, and mounting on slides. The goal is to prepare tissue for microscopic analysis while maintaining structure.
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This document discusses the process of decalcification, which is the removal of calcium salts from bones and calcified tissues. There are four main methods: 1) using simple dilute mineral acids like nitric acid or formic acid; 2) ion exchange resins with acid fluids; 3) electrolytic decalcification using electrodes; and 4) chelating agents like EDTA that bind calcium ions. The document provides details on procedures and advantages/disadvantages of each method.
Histopathology
This document outlines the steps for histology lab procedures which include fixation, processing, sectioning, staining, and mounting of tissues. It lists the required chemicals, glassware, and instruments. The key steps are:
1. Tissues are fixed in formalin or Bouin's fixative for 24 hours to preserve cells and structures.
2. Processing involves dehydration using graded alcohols, clearing with benzene, and infiltration with paraffin wax for embedding.
3. Sections are cut from the wax blocks using a microtome, mounted on slides, and stained with H&E stain.
4. Stained sections are viewed under a microscope after dewaxing and mounting.
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Fixation of special substances
- 3. • In fields of , and fixation is a chemical process by which biological tissues are preserved from decay,thereby preventing from • Fixation terminates ongoing biochemical reactions • It also increase the mechanical strenght or stability of treated tissues.
- 4. • • • • • • • Which are to be fixed are following Proteins Lipids Carbohydrates Biogenic amines Nucleic acids Water and lipid soluble substances
- 5. • Crosslinking fixatives act by creating covalent chemical bonds between proteins in tissue. • Most commonly used fixative in histology is the crosslinking fixative formaldehyde (also named formalin). • Formaldehyde is the most commonly used fixative for amino acid histochemistry. • Another popular aldehyde for fixation is glutaraldehyde. • Glutaraldehyde fix proteins very rapidly. • Glutaraldehyde may not penetrate thicker tissue specimens as effectively as formaldehyde. • Glutaraldehyde may offer a more rigid or tightly linked fixation.
- 6. • In order to demonstrate lipids,frozen sections are commonly used. used to preserve phospholipids. • For preservation of lipids in cryostat sections fixative should contain • For this purpose we use the • Fixation is over 3 days at room temperature.
- 8. • Alcoholic fixatives are used for glycogen fixation. are • Fixatives for ultrasturctural preservation of dextrans and glycogen is • • • • • It is used by making stock solutions of Formaldehyde Glutaraldehyde Osmium tetraoxide Lead citrate
- 9. • Biogenic amines can be demonstrated in tissues using a variety of techniques. • In this reaction potassium dichromateformaldehyde is used. Formaldehyde fixation of the adrenal medulla produced fluorescence of cells which contained biogenic amines or carnoy fixed paraffin section For histamines • Primary biogenic amines are well preserved by
- 10. • Fixatives choosen for nucleic acids must depend upon subsequent methods which are to be employed in analysis of tissue. –based fixatives (used but mostly avoided) – (most widely used) • It is to be used then the hydrolysis time varies with the fixative.
- 11. • For accurate demonstration of water soluble substances,non-aqueous fixatives are used. For example, Urates in gout is a satisfactory fixative For steriods.
- 12. Target Fixatives of choice Fixatives of avoided proteins Natural buffered formalin, paraldehyde Osmium tetroxide lipids Frozen sections Chemical fixative Nucleic acid Alcoholic fixative Aldehyde fixative Glycogen Alcoholic based fixatives Osmium tetroxide