Paraffin Embedding Overview Paraffin Embedding involves the steps fixation, dehydration, transparentizing, immersion and embedding. Paraffin Embedded Tissue is sliced by a rotary microtome to give a thickness of 2-7 Îźm. Paraffin embedding is one of the standard procedures used in preparing slender sections of biological substance for histological assessment by light microscopy. There are major steps that are involved in paraffin embedding includes fixation, dehydration, transparentizing, immersion and embedding. The tissues samples collected are usually submitted to regular pathologic analysis and then preceded through fixation and paraffin embedding. Fixation procedure is used to cross-link the antigens to wellbuilt macromolecules, thereby immobilizing them in tissue. Furthermore, paraffin embedding may have an effect on the immunoreactivity. For predictable histology, fixation of tissue, followed by paraffin embedding is the most widely-used method. It is criterion in the majority of histology laboratories, and is appropriate for assessing lymphoid tissue. Here are the 5 steps described that are involved in paraffin embedding: Fixation The main purpose of this process is to keep cell sharp and tissue in proper shape to prevent putridness, postmortem autolysis, endogenic and exogenic enzyme activity. Maintain the cell structure as well as position by preventing antigen diffusion. It gives color to clarify tissues by diverse affinity to coloring agent. Dehydration In this step the water is completely removed and creates a situation for the next step and hardens the tissue of curiosity. The dehydrating agents Ethanol and Acetone are completely miscible with water and can be arranged in different volumetric ratios. Transparentizing The tissue of interest necessitates a transparentizing move for the reason that the dehydrating agent used in the above step is immiscible with the paraffin. The adding up of transparent reagent helps paraffin soak up into the tissue.
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