Quantification of signal quality and comparison of different hybridization times and probe concentrations.

<p>A. Schematic depicting the manner in which we quantify signal quality via threshold sensitivity. B. Sensitivity of threshold measured in varying probe concentrations and hybridization times. The dotted line represents the sensitivity of a traditional overnight RNA FISH. Error bars reflect standard error of the mean. C. Spot counts for the same conditions as in B. Error bars reflect standard deviation. At 10 minutes and for all probe concentrations, the spot counts for Turbo FISH are statistically different from overnight FISH (4X: p = 9.87×10⁻⁶, 1X: p = 0.0136, 1/4X: p = 4.86×10⁻⁶, 1/16X: p = 1.75×10⁻¹¹; two-tailed t-test). For all conditions, we analyzed spot counts and calculated the sensitivity on 80–120 cells. Data shown represents one of two replicate experiments.