Decision guides

Choose the part of the protocol you need to adjust.

Choose a guide to compare preparation methods, plan controls, select fluorescence channels, or investigate an unexpected staining pattern.

Compare fixation methods by target localization, morphology, and antibody performance.

Choose permeabilization according to target location and the structures that must remain intact.

Compare combined paraformaldehyde and Triton X-100 exposure with a sequential workflow.

Choose retrieval according to specimen preparation, antibody performance, morphology, and background.

Choose controls for specificity, background, target dependence, and imaging artifacts.

Match fluorophores and channels to target abundance, specimen background, and the microscope.

Start from the observed staining pattern, check the relevant controls, and change one factor at a time.