Immunohistochemistry Embedding Techniques
This collection of images showcases various steps involved in embedding processes for immunohistochemistry, highlighting methods like paraffin embedding, tissue sectioning, and staining. Detailed visuals depict the equipment used, such as embedding molds, metal braces, and paraffin ovens. Step-by-step procedures for blocking out and reception of specimens are illustrated, offering insights into the intricate art of preparing tissue samples for analysis.
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Presentation Transcript
University of TISHK Immunohistochemistry Embedding Dr. Ahmad H. Ibrahim 10 Email: ahmad.ibrahim@Tiu.edu.iq 1
Paraffin method Embedding Blocking out
MICROTECHNIQUES Tissue Cytology Histopathology Body fluids/ Smears from Specimen Biopsy / Autopsy aspiration or exfoliation Method Paraffin / Frozen Smears / Imprint
Reception of specimen Gross pathology Fixation Processing Cover slip Embedding Cutting with Microtome Staining Mounting
Principle The processed tissue section is placed inside special molds to make solid paraffin block which fix and hold the tissue in well oriented position in order to get thin sections by microtome.
Embedding can be done by: Manual Special machine (Embedding station)
Generally we need the following parts or places: Paraffin oven Hot plate Cold plate Embedding molds Paraffin oven: a hot container to keep the melted paraffin inside it which is connected to a drain to use it.
Molds Metal molds L-shaped brace
Blocking is carried out as follows: 1. Adjust the mold: to suitable size in case of L-shaped brace, or appropriate size of metal type, and then fill with melted wax.
Blocking is carried out as follows: 2. Take the tissue container out of the oven, open the cassettes and hold the tissue gently with a warmed forceps.
Blocking is carried out as follows: 3. put it down the melted wax-filled mold
Blocking is carried out as follows: 4. Wait until the lower layer of paraffin solidify. File:Tissue processing - Embedding station.jpg
Blocking is carried out as follows: 5. Gently press the tissue down as far as it will go in the wax and allow it to harden. File:Tissue processing - Embedding station.jpg
Blocking is carried out as follows: 6. fill the remaining of the mold with paraffin to make paraffin block, and place the label over it, either a paper or labeled tissue cassettes.
Blocking is carried out as follows: 7. Put it on a cold plate to become harden and allow to remove the paraffin block contain the tissue, which will be ready for further steps. File:Tissue processing - Moulds are set on a cold plate to solidify.jpg
File:Tissue processing - Solidified paraffin block is popped out of the metal mould.jpg
File:Tissue processing - After metal mould is removed, tissue can be seen in the paraffin block.jpg
Orientation of specimens: The tissue must be putted down the paraffin block according to the following points: 1. The cut surface or flat surface. 2. The layers of tissue or organs. 3. The surface that not stained by ink.
File:Tissue processing - After metal mould is removed, tissue can be seen in the paraffin block.jpg File:Tissue processing - Solidified paraffin block is popped out of the metal mould.jpg File:Tissue processing - Embedding station.jpg File:Tissue processing - Moulds are set on a cold plate to solidify.jpg
