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==Procedures== *Bacteria cells washing and adjusting **Pallet by 4000 rpm, 5 min **Resuspended at 0.5 mL PBS **Adjust to OD600 = 1.0 (10^6 CFU/uL) using Nanodrop *Fixation (Start with 200 uL diluted (10^6 CFU/uL)E.coli (2x10^8 cells) in a 1.5-mL tube) **Pellet the bacteria by 4000 rpm, 5 min **Resuspended the bacteria in 100% ethanol. **Incubate at RT on rocking plate (speed=3) for 3 hr **All fixed cell are stored in -20C *Hybridization **Pre-warmed the hybridization buffer (200 uL) and washing buffer (500 uL) at 46-48C **The fixed cells (10^8) were washed with 0.5 mL PBS twice and resuspended in 200 uL of hybridization buffer. Hybridize for 1.5 hr at 46C, 300 rpm using eppendorf isothermo mixer. **Pellet the labeled bacteria by 4000 rpm, 5 min. Remove the supernatant. Resuspended in 500 uL washing buffer and incubate at 48C, 300 rpm for 20 min. **Pellet the labeled bacteria by 4000 rpm, 5 min. Remove the supernatant. Resuspended in 1 mL cold PBS. Wrapped in foil and stored in 4C.
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