Materials and Reagents

1. Adherent-invasive E. coli reference strain LF82 (Darfeuille-Michaud et al., 1998)
   
2. Intestine-407 (I-407) cells (ATCC, catalog number: CCL-6 )
   
3. Luria Broth (BD Difco^TM, catalog number: 244620 )
   
4. Glucose (Sigma-Aldrich, catalog number: G8270 )
   
5. M63 medium (United States Biological, catalog number: M1015 )
   
6. MgSO₄ (Sigma-Aldrich, catalog number: 208094 )
   
7. Phosphate-Buffered Saline (PBS) ( Mediatech, Cellgro^®, catalog number: 21-040 )
   
8. Crystal Violet (Sigma-Aldrich, catalog number: C6158 )
   
9. 200 proof absolute Ethanol (Sigma-Aldrich)
   
10. Fetal bovine serum (FBS) (Mediatech, Cellgro^®, catalog number: 35-010-CV )
    
11. Nonessential amino acids (Mediatech, Cellgro^®, catalog number: 25-025-CI )
    
12. L-glutamine (Mediatech, Cellgro^®, catalog number: 25-005 )
    
13. Penicillin/Streptomycin/Amphotericin B solution (Mediatech, Cellgro^®, catalog number: 30-004-CI )
    
14. Vitamin mix (Mediatech, Cellgro^®, catalog number: 25-020-CI )
    
15. Formaldehyde solution (Sigma-Aldrich, catalog number: F8775 )
    
16. Phalloidin-tetramethyl rhodamine isocyanate (Sigma-Aldrich, catalog number: P1951 )
    
17. Paraformaldehyde
    
18. Hoechst 33258 (Sigma-Aldrich, catalog number: B1155 )
    
19. Vectashield (Vector Labs, catalog number: H-1000 )
    
20. Minimum Essential Medium (MEM) (Mediatech, Cellgro^®, catalog number: 10-022-CV ) (see Recipes)
    

Equipment

1. Coverslips (Electron Microscopy Sciences)
   
2. Sterile tubes
   
3. 24-well tissue culture plates
   
4. Non-cell-treated polystyrene 96-well microtiter plates (Falcon^®, catalog number: 62406-117 )
   
5. 24-well polystyrene plate, tissue-culture treated (Falcon^®, catalog number: 62406-159 )
   
6. Microplate shaker (LABREPCO, model: BT1500 )
   
7. Microplate spectrophotometer
   
8. Confocal microscope
   
9. 30 °C incubator
   
10. 37 °C/5% CO₂ incubator
    

Software

1. Computer program COMSTAT1

Procedure