TABLE 2.

Comparison of fluorescence intensities and hybridization efficiencies between one-step and two-step FISH and between Eco541 and Eco1482 probes at different temperatures and hybridization times^a

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

^aData 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).

^bIndicates pretreatment of sample at 90°C for 5 min.