Protocol aim
The aim of this protocol is to provide instructions for paraffin embedding of fixed, cell laden constructs. Embedded samples can, among other applications, be stained for immunofluorescence and immunohistology analysis. Follow Fixation Protocol before starting this protocol.
Materials Needed
- Paraffin
- Dry oven at 58°C
- Fixed, cell laden construct
- Embedding cassettes
- 70% Ethanol
- 96% Ethanol
- 100% Ethanol
- Xylene or xylene substitute, e.g. Shandon Xylene Substitute (Thermofisher, Ref: 9990505)
- Tissue embedding machine
Protocol
This protocol can be performed non-sterile, note that all handling and use of ethanol and xylene/xylene substitute must be done inside a fume hood with proper PPE. Dispose waste according to local regulations. If performing dehydration and paraffin infiltration with tissue dehydration and infiltration machine, it’s recommended to test the automated process with spare samples before using with sensitive samples. If dehydration and paraffin infiltration is done with a tissue dehydration and infiltration machine only step 2 and 5 is performed in the protocol.