The method of application varies depending on the quenching agent and the type of tissue. Generally, tissue sections are treated with a solution containing the quenching agent after fixation but before staining. For instance, a common protocol involves immersing tissue sections in a 0.1% solution of Sudan Black B in 70% ethanol for 10-15 minutes, followed by rinsing in ethanol and water. The specifics of concentration, incubation time, and washing steps can vary and should be optimized for each experimental setup.
