Skip to main content
. 2018 May;18(2):132–140. doi: 10.17305/bjbms.2018.2519

FIGURE 3.

FIGURE 3

Examples of fluorescence in situ hybridization (FISH) assays that were inadequate for scoring. (A) Increased binding of DAPI, autofluorescence, and absent signals in nuclei due to incomplete hybridization and shorter digestion, Vysis/Abbott pretreatment kit with 10-minute protease digestion (DAPI counterstain, ×1000); (B) Disrupted nuclear morphology, inadequate DAPI binding and loss of signals due to prolonged pepsin digestion, DAKO pretreatment kit with 15-minute pepsin digestion (DAPI counterstain, ×1000); (C) Loss of signals due to prolonged post-hybridization washing with higher temperature, DAKO pretreatment kit, 20×Tris/HCl, 75°C/10 minutes (DAPI counterstain, ×1000); (D) Excessive, non-specific signals due to non-stringent wash, (Spectrum green, ×1000); (E) Background autofluorescence masking the signals, Vysis/Abbott pretreatment kit, 2×SSC/0.3% NP-40, 60°C/5 min (Spectrum Green, ×1000); (F) Unsuccessful hybridization in case of prolonged fixation time, using Vysis/Abbott pretreatment kit (DAPI counterstain, ×1000). SSC: Saline sodium citrate; NP40: Nonionic detergent; DAPI: 4’,6-diamidino-2-phenylindole.