Once the staining solutions and associated reagents have been selected, it is good to start with the baseline H&E staining protocol. From there, edit either the hematoxylin in 30-second increments OR the eosin in 15-second increments. Remember, eosin will tend to penetrate much faster. Unless there is the need to significantly lighten or darken the eosin staining intensity, the shorter increments are best. It is also important that only one stain is changed at a time. It may appear that the hematoxylin is overstained, but in reality, the eosin just needs to be richer.
