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Protocols Used at NCI and Related Information, Paraffin Embedding

 
Paraffin Embedding

This method was successful in our lab using prostate tissue and for our specific objectives. Investigators must be aware that they will need to tailor the following protocol for their own research objectives and tissue under study.

1. Materials

  1. Sakura FineTek V.I.P. tissue processor (Sakura FineTek Inc.)
  2. 70%, 80%, 95%, 100% ethanol
  3. Paraffin wax (Paraplast; melting point 58°C)
  4. Xylenes, Reagent Grade (Sigma)
  5. Embedding molds

2. Method

  1. Fix tissue overnight in 70% EtOH at 4°C.
  2. Place the tissue in the tissue processor and process with the times and temperatures described in Table 1 below.
  3. Embed the specimen in paraffin and block.
  4. Cut sections at 5 to 8 µm thickness.
  5. Both paraffin blocks and sections may be stored at or below room temperature.

    TIP: Tissue processors in standard histopathology laboratories generally include formalin-fixation as the first two steps in the paraffin infiltration procedure. Make certain these steps are avoided when processing tissue intended for molecular profiling.

Table 1

Station

Solution

Concentration

Time(min)

Temp(°C)

1 Ethanol 70% 30 40
2 Ethanol 80% 30 40
3 Ethanol 95% 45 40
4 Ethanol 95% 45 40
5 Ethanol 100% 45 40
6 Ethanol 100% 45 40
7 Xylenes 100% 45 40
8 Xylenes 100% 45 40
9 Paraffin   30 58
10 Paraffin   30 58
11 Paraffin   30 58
12 Paraffin   30 58

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