石蜡加填充剂 Paraffin Embedding

互联网2013-12-26

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Paraffin Embedding 石蜡加填充剂

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

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

2. Method

Fix tissue overnight in 70% EtOH at 4°C.
Place the tissue in the tissue processor and process with the times and temperatures described in Table 1 below.
Embed the specimen in paraffin and block.
Cut sections at 5 to 8 µm thickness.
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.

<center> <blockquote> </blockquote> Table 1 <table> <caption> </caption> <tbody> <tr> <th> Station </th> <th> Solution </th> <th> Concentration </th> <th> Time(min) </th> <th> Temp(°C) </th> </tr> <tr> <td> 1 </td> <td> Ethanol </td> <td> 70% </td> <td> 30 </td> <td> 40 </td> </tr> <tr> <td> 2 </td> <td> Ethanol </td> <td> 80% </td> <td> 30 </td> <td> 40 </td> </tr> <tr> <td> 3 </td> <td> Ethanol </td> <td> 95% </td> <td> 45 </td> <td> 40 </td> </tr> <tr> <td> 4 </td> <td> Ethanol </td> <td> 95% </td> <td> 45 </td> <td> 40 </td> </tr> <tr> <td> 5 </td> <td> Ethanol </td> <td> 100% </td> <td> 45 </td> <td> 40 </td> </tr> <tr> <td> 6 </td> <td> Ethanol </td> <td> 100% </td> <td> 45 </td> <td> 40 </td> </tr> <tr> <td> 7 </td> <td> Xylenes </td> <td> 100% </td> <td> 45 </td> <td> 40 </td> </tr> <tr> <td> 8 </td> <td> Xylenes </td> <td> 100% </td> <td> 45 </td> <td> 40 </td> </tr> <tr> <td> 9 </td> <td> Paraffin </td> <td> </td> <td> 30 </td> <td> 58 </td> </tr> <tr> <td> 10 </td> <td> Paraffin </td> <td> </td> <td> 30 </td> <td> 58 </td> </tr> <tr> <td> 11 </td> <td> Paraffin </td> <td> </td> <td> 30 </td> <td> 58 </td> </tr> <tr> <td> 12 </td> <td> Paraffin </td> <td> </td> <td> 30 </td> <td> 58 </td> </tr> </tbody> </table> </center> <center> </center>
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