In some IHC detection reagent instructions, the last step states “dehydrate, clear and mount”. What does this mean?
In a manual IHC application, these steps involve submersion of the slides into a graded ethanol series, immediately followed by submersion into a miscible solution (xylene or alternative), followed by placement of a coverslip over the preparation with a suitable mounting medium. These steps preserve the specimen for subsequent visualization, imaging and archiving. From our experience, we suggest moving the slides in a slide rack through a series of dishes, commencing dehydration at 95% ethanol (2 changes for 2 min ea.), then into 100% ethanol (2x 2 min ea.), then into clearing solution (3x 2 min ea.), followed by mounting (coverslipping).