There are various protocols for using toluidine blue, depending on the specific application. A general staining protocol may include the following steps:
Preparation of the tissue section: The tissue is fixed and embedded in a suitable medium, such as paraffin or resin. Thin sections are then cut using a microtome. Hydration: The sections are hydrated through a series of alcohol and water washes to prepare them for staining. Staining: The sections are immersed in a toluidine blue solution for a specified duration, typically ranging from a few seconds to several minutes. Rinsing: Excess dye is removed by rinsing the sections in distilled water or a buffer solution. Dehydration and mounting: The sections are dehydrated through a series of alcohol washes, cleared in xylene, and mounted with a suitable mounting medium.
