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When sperm with their plasma membrane permeabilized with SLO are exposed to Fluo3-AM, the dye diffuses into the acrosome, where esterases remove the AM moiety, trapping the sensor in this compartment.
Thanks to the SLO permeabilization protocol, we could actually visualize the human sperm acrosomal calcium store with Fluo3-AM, something not achievable in non-permeabilized sperm, where cytosolic esterases remove the AM moiety, trapping membrane impermeant Fluo3 in the cytosol.
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