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Basic Principle of IHC Fixation and Staining - Immunostaining

IHC fixation method for tissue sample. This IHC fixation protocol provides the detailed descriptions paraformaldehyde, ethanol, methanol, and acetone fixation methods. Optimize your immersion time by reading involves IHC with similar tissue types as yours.

Immunostaining
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Basic Principle of IHC Fixation and Staining - Immunostaining

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  1. What Is Fixation???? Website : www. immunostaining.info

  2. Contents • Introduction • Purpose • Effect of Fixation • Summary

  3. Introduction Preparation of tissue for microscopic examination , through a series of namely fixation , clearing, embedding ,cutting & Staining. Tissue decompose when removed from the body or it is cut off from the body supply – Autolysis or Putrefaction

  4. Definition of Fixation • A physical or chemical process that stops as soon as possible vital cellular processes, maintaining with minimal alternation the shape, volume spatial and molecular relation between elements. • Fixation is the foundation for the subsequent stages in the preparation of the sections.

  5. Purpose 1.To prevent autolysis , bacterial decomposition & putrefaction. 2. To coagulate the tissue as prevent loss of easily diffusible substances. 3. Indurate tissues to enhance working with glass slides. 4. To facilitate differential staining with dyes & other reagents.

  6. Effect of Fixation • Inhibitions of autolysis & putrefaction • Preservation • Hardening • Solidification of colloid material • Optical differentiation • Effect on staining

  7. Method of Fixation • Immersion • Perfusion • Perfusion – immersion • Heat fixation

  8. Summary • It is clear that fixation is normally carried out as soon as possible to prevent autolysis and to reduce possible infectivity. • Formalin is used for all routine surgical pathology and autopsy tissues when an H & E slides is to be produced.

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