Tissue processing

tissue processing of histopathological tissue

1. Tissue processing Presented by Aniket choudhary

2. DEFINATION OF TISSUE PROCESSING Tissue processing is the technique by which fixed tissue are made suitable for embedding within a supportive medium such as paraffin.

3. TISSUE PROCESSING CONSIST OF SOME STEPS :- FIXATION DEHYDRATION CLEARING INFILTRATION EMBEDDING BLOCK MAKING SECTION CUTTING ROUTINE STAINING MOUNTING

4. FIXATION Fixation is a chemical process by which biological tissue are preserved and hardening of the organ in such a way that its original form is retain and its constituents do not spread out. Commonly used fixative in lab 10% formalin Formal saline 10% buffered formaline Alcoholic formaldehyde

5. 2. DEHYDRATION Dehydration is a process of removal of water from tissue. Alcoholic remove unbound water from tissue sent to a alcohol 70%, 80%, 95%, ethanol , methyl and action can be used.

6. 3. CLEARING Zylene is commonly used for clearing a tissue. Zylene is helpful in removal of alcohol it fixed tissue. Zylene , toluene, benzene, chloroform can be as a clearing agent .

7. 4. INFILTRATION Infiltration is the process why which clearing agent is completely remove from the tissue. Clearing agent is estimated from the tissue by defusion in the surrounding. The process when be added the melted paraffin wax in tissue is called the infiltration .

8. 5. EMBEDDING The principal of embedding is to accurately and presively oriented a histological sample of the tissue into a block of paraffin wax. To L-mould are used for making a block will allow the support and for to hold the tissue so that find cutting and thick section will be cut easily in a microtomes.

9. 6. BLOCK MAKING Metals L-moulds smeared with paraffin wax Orient sample so that intend the cutting surface is preserved against base. Fill the moulds with fresh metal wax. Place identifying label. When block completely solidity remove mould. Block ready for cutting.

10. 7. Section cutting Section cutting by using the microtome:- Fix the block holder of the microtome. Insert tightly a knife in the knife holder with proper position. Cut the sufficient section.

11. 8. ROUTINE STAINING “H” and “E” staining can be used as a routine stain. “H” and “E” means Hematoxyline and Eosin.

12. 9. MOUNTING Mounting by using DPX. Help to keep a sample in plays during imagine. Prevent the specimen from drying out. Preserve sample over time for long term storage.

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