Fluorescent FM dyes partition reversibly into biological membranes without penetrating through them. Suppose that you have neurons cultured in a dish. You incubate the neurons with an FM dye, and then you wash them with medium to remove the dye. You find that if the neurons are stimulated to trigger action potentials during incubation with the dye, the synaptic regions of the neurons remain fluorescent after the wash. Can you explain why this effect may have occurred? Also, after generating fluorescent synaptic regions by this procedure, suppose that you trigger additional action potentials while continuing to wash with medium. Would you expect the synaptic regions to lose fluorescence?

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