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. 2012 Jul 24;7(7):e40675. doi: 10.1371/journal.pone.0040675

Figure 2. Effect of EC on hybridization rate.

Figure 2

Time chase comparison of FISH signal intensities using 15% EC buffer and 45% formamide buffer on FFPE breast carcinoma tissue sections. Cot-1 blocking was used in formamide buffer, but omitted for EC buffer. Identical probe concentration was used in the two buffers. EC buffer was denatured at 67°C for 10 minutes, formamide buffer at 82°C for 5 minutes, and then both hybridized at 45°C from t = 0 minute to t = 23 hours. A: Merged micrograph of red HER2 DNA probe signals, green CEN-17 PNA signals and blue DAPI staining (Figure S2). The images are taken with identical exposure times. Scale bar, 10 µm. B: Quantitative analysis of HER2 DNA and C, of CEN-17 PNA signals (Figures S2 and S3 ). MFI, mean fluorescence intensity. The bars represent the 95% confidence interval (n = 45 signals).