Materials and Reagents

1. M. mobile 163K strain
   
2. M. pneumoniae M129 strain
   
3. Horse serum (Life Technologies, catalog number: 16050-122 )
   
4. Interested reagents (for example, 0.05-1 mM 3’-sialyllactose, Dextra Laboratories, catalog number: SL302 )
   
5. Heart infusion broth (Becton, Dickinson and Company, catalog number: 238400 )
   
6. Yeast extract (Becton, Dickinson and Company, catalog number: 212750 )
   
7. Amphotericin B (Sigma-Aldrich, catalog number: A2942 )
   
8. Ampicillin (Nacalai Tesque, catalog number: 02739-32 )
9. NaCl
   
10. Sodium phosphate (pH 7.3)
    
11. Glucose
    
12. Aluotto medium (see Recipes)
    
13. PBS-G buffer (see Recipes)
    

Equipment

1. Test tubes
   
2. Tissue culture flask
   
3. Scraper (Greiner Bio-One GmbH, catalog number: 541070 )
   
4. 0.45-μm-pore size filter (Millipore, catalog number: SLHVX13NK ) (PolyVinylidene DiFluoride, or equivalent)
   
5. 25 °C incubator (Yamato, model: IL62 )
   
6. 37 °C incubator (Yamato, model: IS62 )
   
7. Centrifuges (Sigma Zentrifugen, model: SIGMA 1-14 )
   
8. Optical microscope (OLYMPUS, model: BX51 )
   
9. Stage heater (for M. pneumoniae) (Minitube, model: HT200 )
   
10. Lens heater (for M. pneumoniae) (Tokai Hit, model: MATS-LH )
    

Software

1. ImageJ

Procedure