Comparison of fixation conditions for both traditional overnight hybridizations and rapid hybridization.
A. Comparison of number of spots detected and cumulative distribution functions for the TBCB gene with probes labeled with the Alexa 594 fluorophore. Error bars represent the standard error of the mean. No statistically significant differences exist between the overnight RNA FISH samples. Turbo RNA FISH for TBCB gene on formaldehyde-fixed cells is statistically different from Turbo RNA FISH on methanol- and ethanol-fixed cells (p = 3.82×10−65 and p = 4.89×10−96, respectively; two-tailed t-test). For all conditions, we analyzed spot counts on 100–150 cells. B. Comparison of number of spots detected and cumulative distribution functions for the TOP2A gene with probes labeled with the Cy3 fluorophore. Error bars represent the standard error of the mean. Overnight RNA FISH for TOP2A gene on formaldehyde-fixed cells is statistically different from overnight RNA FISH on ethanol-fixed cells (p = 0.0067; two tailed t-test). No other statistically significant differences exist between overnight RNA FISH samples. Turbo RNA FISH for TOP2A gene on formaldehyde-fixed cells is statistically different from Turbo RNA FISH on methanol- and ethanol-fixed cells (p = 9.57×10−28 and p = 4.22×10−30, respectively; two-tailed t-test). For all conditions, we analyzed spot counts on 100–150 cells. Data shown represents one of two replicate experiments.
