RNA CISH Signal Detection: A Technique to Detect Chromogenic Signals During RNA In Situ Hybridization in Intact Tissue Specimens

Dispense the same number of drops of each DAB-A and DAB-B solution in an appropriately sized tube to make approximately 120 microliters of DAB substrate per section and vortex. Take each slide, one at a time, from the slide rack and tap and flick to remove excess liquid, and place it back in the slide rack. Pipette approximately 120 microliters of DAB mixture onto each tissue section, making sure that the sections are covered. Incubate for 10 minutes at room temperature and proceed with counterstaining.

Place one drop of mounting medium per glass slat and then place the slat on the slides and let them air dry. After a few hours, proceed with evaluation using an optical microscope as described in the manuscript.