Fig. 4: AmpFISH is adopted to robustly assay transcripts in mouse brain slices.
From: Preparation of long single-strand DNA concatemers for high-level fluorescence in situ hybridization
a Schematic diagram depicting four different experimental workflows. b FISH imaging of Gad1 gene in mouse brain slices using different pipelines depicted in (a) (scale bar: 10 μm). c Statistical analysis of signal-to-noise ratio (SNR) of AmpFISH from four experimental pipelines in (b). The intensity of an area as background signal next to the signal was subtracted from the signal itself. The signal-to-noise ratio was determined by formula: (Signal-background)/background. One-way analysis of variance (ANOVA), F(3, 72) = 10.83, P < 0.0001; Bonferroni’s multiple comparison test, no treatment vs. 0.2% SDS, t = 4.206, P < 0.001; no treatment vs. 1% SDS, t = 4.810, P < 0.001; methanol vs. 0.2% SDS, t = 3.030, P < 0.05; methanol vs. 1% SDS, t = 3.633, P < 0.01. (n = 19, from three repeat experiments). d The specificity of AmpFISH via secondary amplification hybridization was verified in brain slices by expressing EYFP protein in Thy-EGFP transgenic mouse (scale bar: 25 μm).
