Materials and Reagents

1. Adherent tumor cells [such as non-small cell lung carcinoma (NSCLC) cell lines]
   
2. Specific T-cell clones (generated either from autologous tumor-infiltrating T lymphocytes (TIL) or peripheral blood lymphocytes (PBL))
   
3. RPMI 1640 (Life Technologies, Gibco^®, catalog number: 61870044 )
   
4. DMEM-F12 (Life Technologies, Gibco^®, catalog number: 31331093 )
   
5. UltroserG (Pall, catalog number: 15950-017 )
   
6. Fetal Bovin Serum (Life Technologies, Gibco^®, catalog number: 10270-106 )
   
7. Human serum AB (Institut de Biotechnologies Jacques Boy)
   
8. Penicillin and streptomycin (Life Technologies, Gibco^®, catalog number: 15140122 )
   
9. Sodium Pyruvate (Life Technologies, Gibco^®, catalog number 11360029 )
   
10. IL-2
    
11. 10x PBS (Life Technologies, Gibco^®, catalog number: 70011-036 )
    
12. CellTracker probe (CellTracker^TM Green CMFDA) (Life Technologies, Invitrogen™, catalog number: C2925 )
    
13. Complete DMEM: tumor cell culture medium (LC medium) (see Recipes)
    
14. RPMI-based T-cell complete medium (see Recipes)
    

Equipment

1. Microscope Zeiss LSM-510 (ZEISS) with a heated incubation chamber and CO₂ supply
   
2. Micro-Slides VI, ibiTreat (ibidi GmbH, catalog number: 80606 ), two silicon tubes (1.6 mm of inner diameter) with a plastic clip, two Elbow Luer connectors (ibidi GmbH, catalog number: 80646 )
   
3. Syringe pump (high flow rate > 50 ml/min)
   
4. 60 ml syringe (Becton, Dickinson and Company, catalog number: 300866 )
   
5. Humidified incubator at 37 °C with 5% CO₂
   
6. A recipient for waste flow buffer (Erlenmeyer)
   
7. Centrifuge (Beckman Coulter, model: GS-6R )
   

Procedure