Protocols
Formalin fixed-paraffin-embedded cell line pellet preparation
Protocol created by Scherezade Jiménez and Patricia González - Monoclonal Antibodies Unit and Histopathology Unit, Centro Nacional de Investigaciones Oncológicas
Matherial and reagents:
- Formalin solution neutral buffered (HT501128-4l Sigma)
- Tissue-Tek Paraform cassette frames, white. (Ref.7040)
- Tissue-Tek Paraform Biopsy cassettes, 13x13. (Ref.7019)
- Phosphate buffered saline, (PBS) (79378 Fluka Analytical)
- Before preparing the pellet verify the status of the cell culture (at least 28x106 cells are needed).
- Wash your pellet with PBS (5 minutes 1000prm).
- Resuspend you pellet in 2ml PBS and place it in a 2ml eppendorf. Centrifuge 5 min a 1200rpm.
- Resuspend the pellet in 2ml Eppendorf with 1 ml of formalin (10%) and mix gently (keep it in movement at room temperature for 1 hour).
- Centrifuge at 13000rpm for 5 minutes.
- Discard the formalin and collect the pellet using a metal spatula.
- Place the pellet in a special cassette (Tissue-Tek Paraform cassette, see image below)
- Placed the cassette in a tissue processor.
