TABLE 2.
FISH conditions | Fluorescence intensity | SD | FISH efficiency (%) | SD |
---|---|---|---|---|
Std | 31.6 | 1.9 | 83.9 | 9.0 |
46°C, 30 min | 27.8 | 10.4 | 88.7 | 7.9 |
55°C, 30 min | 59.2 | 4.5 | 87.5 | 4.3 |
60°C, 30 min | 59.2 | 26.2 | 95.7 | 8.3 |
65°C, 30 min | 68.3 | 5.6 | 106.2 | 19.5 |
50°C, 20 minb | 73.7 | 3.1 | 100.1 | 6.3 |
SYBR Green | 37.9 | 3.1 | 100.0 | 8.8 |
Eco541-Std | 203.6 | 5.1 | 58.7 | 1.6 |
Eco1482-Std | 159.0 | 1.5 | 66.1 | 5.1 |
Eco541-55°C, 30 min | 199.7 | 0.8 | 82.6 | 4.7 |
Eco1482-55°C, 30 min | 150.6 | 0.4 | 105.5 | 2.7 |
Data are also illustrated in Fig. 6 and 7. The fluorescence intensities of hybridized E. coli cells were normalized to 0.75-μm beads, while the hybridization efficiencies were normalized to the cell counts for the standard conditions (Std) (46°C; 3 h) (for comparison of one-step and two-step FISH) or normalized to the counts of reference beads (for comparison of the two probes).
Indicates pretreatment of sample at 90°C for 5 min.