- Nuclear: The staining times, the need for a differentiation step or not, and the bluing method will be dictated by which type of hematoxylin is to be used. The next page lists some commonly used hematoxylins and their staining times.
- Cytoplasmic: The intensity of eosin staining and the degree of differentiation is mainly a matter of individual preference. Therefore, the type of eosin used, the concentration of the eosin, the staining time and the length of time in the subsequent dehydrating alcohols, can all be optimized for the individuals' satisfaction.
4. Dehydration: In addition to differentiating the eosin, the purpose of the graded alcohols is to remove the water from the tissue sections. This final step is necessary to prepare the sections for coverslipping in a non-aqueous mounting media.
5. Clearing: After several changes of the clearing agent, the slides are ready to be coverslipped and viewed under the microscope.