To achieve effective clearing, follow these steps: 1. Dehydrate the Tissue: Ensure that the tissue is thoroughly dehydrated using a graded series of alcohols, typically finishing with 100% ethanol. 2. Immerse in Clearing Agent: Transfer the tissue to the clearing agent. The duration depends on the tissue type and the clearing agent used. For example, xylene generally requires 1-2 hours. 3. Monitor Transparency: Check the tissue periodically for transparency. The tissue should become clear, indicating that it is ready for the embedding process. 4. Multiple Changes: For thicker or denser tissues, multiple changes of the clearing agent may be necessary to ensure complete clearing.
